LOCUS       Exported                7099 bp ds-DNA     circular SYN 10-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7099)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Jul 10, 2021 from SnapGene Viewer 5.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7099
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2234..2268
                     /label=source
     source          join(5227..7099,1..2233)
                     /label=source
     rep_origin      complement(145..733)
                     /direction=LEFT
                     /label=ori
     primer_bind     complement(225..244)
                     /label=pBR322ori-F
     CDS             complement(904..1764)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1527..1546
                     /label=Amp-R
     promoter        complement(1765..1869)
                     /label=AmpR promoter
     primer_bind     1937..1955
                     /label=pBRforEco
     primer_bind     complement(1993..2015)
                     /label=pGEX 3'
     primer_bind     2115..2134
                     /label=pRS-marker
     promoter        2215..2233
                     /label=SP6 promoter
     primer_bind     2215..2232
                     /label=SP6
     CDS             2274..2981
                     /codon_start=1
                     /label=mCherry
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2962..2981)
                     /label=ase1_pnmt1-mCherry_rvs
     misc_feature    2982..5224
                     /label=ase1 gene
     exon            2982..3405
                     /label=exon
     misc_feature    3406..3452
                     /label=nonstandard type: cds_intron
     exon            3453..5224
                     /label=exon
     misc_feature    5225
                     /label=nonstandard type: three_prime_utr
     terminator      5245..5432
                     /label=ADH1 terminator
     primer_bind     complement(5300..5324)
                     /label=pHybLex-R
     gene            5507..6863
                     /label=kanMX
     promoter        5507..5850
                     /label=TEF promoter
     CDS             5851..6660
                     /codon_start=1
                     /label=KanR
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      6666..6863
                     /label=TEF terminator
     primer_bind     complement(6967..6986)
                     /label=T7
     promoter        complement(6968..6986)
                     /label=T7 promoter
     primer_bind     complement(7073..7090)
                     /label=L4440
ORIGIN
        1 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
       61 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
      361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
      601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
      781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
      841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     1921 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     1981 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2041 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2101 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
     2161 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     2221 gtgacactat agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacatggtga
     2281 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2341 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2401 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2461 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2521 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2581 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2641 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     2701 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     2761 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2821 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2881 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2941 agggccgcca ctccaccggc ggcatggacg agctgtacaa gatgcaaaca gtaatgatgg
     3001 atgacattca aagcactgat tctattgctg aaaaagataa tcactctaat aatgaatcta
     3061 actttacttg gaaagcgttt cgtgaacaag tggaaaagca tttttctaaa attgaaaggc
     3121 ttcaccaagt ccttggaaca gatggagaca attcatcatt atttgagttg tttacaacgg
     3181 caatgaatgc ccagcttcat gaaatggaac agtgccagaa aaaacttgaa gatgactgtc
     3241 agcaaagaat tgattcaatc agatttttgg tttcctcatt aaagttaacg gatgatactt
     3301 ctagtctcaa aattgagtct cctttaattc agtgtttgaa tcgtttgtca atggtagaag
     3361 gacaatatat ggcacagtat gatcaaaagt taagtacgat taaaggtatg taatcgtctt
     3421 taatttagac ttgtgtttta actgatgtat agaaatgtat cacaaattgg agtcatattg
     3481 taaccgctta ggaagtccgt tcgttttacc tgattttgag aattcatttt tatctgatgt
     3541 atccgatgct tttactgaat ctttgagagg acgcatcaac gaagccgaaa aggagattga
     3601 tgcgagatta gaggttatta attcctttga agaagaaatt ttgggtttgt ggtctgaact
     3661 cggtgttgag cccgctgatg ttccacaata cgaacaattg cttgaatccc atactaatcg
     3721 accaaatgat gtttatgtta ctcaagaact tatcgaccaa ctttgcaagc aaaaagaagt
     3781 tttttccgct gaaaaagaaa agagaagtga tcatttaaaa agtatacaat cagaagttag
     3841 caacttgtgg aataagcttc aagtttctcc caatgaacaa agtcaatttg gcgattcatc
     3901 aaacattaat caagaaaata tttcattatg ggaaactgaa cttgaaaaac ttcatcagtt
     3961 aaaaaaggag catttaccca tttttttaga agactgtcgt caacaaattc ttcagctttg
     4021 ggattctctg ttttattcag aagaacaaag aaagtccttt acacctatgt atgaagacat
     4081 tattacagag caggttctta cggcccatga aaactatata aagcaactag aggccgaagt
     4141 ttctgctaat aagtcctttt taagcttaat taatcgctat gcctctttaa tagaaggaaa
     4201 gaaagagctt gaagctagtt ctaatgatgc ctctcgtcta acacaacggg gacgccggga
     4261 cccaggttta cttctacgtg aagagaaaat ccgtaagcga ctttctagag aacttcctaa
     4321 ggttcagtcg ctgcttatac cagagattac agcatgggaa gaaagaaatg gaaggacgtt
     4381 ccttttttat gatgaaccac ttctcaagat ttgccaagag gccactcaac caaaatcatt
     4441 atatagaagt gcaagtgctg ccgcaaaccg cccgaaaaca gcaactacaa cggactctgt
     4501 taatagaaca ccttctcaac gagggcgtgt agctgtacct tcaacaccaa gtgttaggtc
     4561 cgcttctcga gctatgacga gtccaaggac accgcttcct agagtaaaaa acactcaaaa
     4621 tccaagtcgt tccattagtg cagaaccgcc atcagcaacc agtaccgcca atagaagaca
     4681 ccccactgct aatcgaattg atataaacgc tagattaaac agtgctagtc ggtctcgaag
     4741 cgcgaacatg ataagacaag gggcaaatgg tagtgacagc aatatgtctt cttcacccgt
     4801 ttctggaaat tccaataccc cttttaacaa gtttccaaat tctgtatctc gcaatacaca
     4861 ttttgaatcc aagtcaccgc acccaaatta ctctcgaact cctcatgaaa cgtattcaaa
     4921 ggcttcatct aagaacgtcc cattaagtcc tccaaagcag cgtgtagtta atgaacacgc
     4981 tttaaatatt atgtcggaaa aattgcaaag aactaatctg aaagaacaaa cacccgagat
     5041 ggacattgaa aacagctcgc agaaccttcc tttttctcct atgaagatat cccccataag
     5101 agcatcaccc gtaaagacaa ttccatcatc accgtccccc actaccaaca ttttttctgc
     5161 tccactcaac aatattacaa attgtacacc gatggaggat gaatggggag aagaaggctt
     5221 ttaaggcgcg ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     5281 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     5341 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat
     5401 gaggtcgctc ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata
     5461 tagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca
     5521 gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc
     5581 gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg
     5641 cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc
     5701 cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta
     5761 ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag
     5821 ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg
     5881 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     5941 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     6001 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     6061 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     6121 gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa
     6181 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     6241 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     6301 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     6361 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
     6421 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     6481 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     6541 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     6601 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     6661 tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     6721 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     6781 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     6841 tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa
     6901 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     6961 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     7021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7081 ctgactcgct gcgctcggt
//
