LOCUS       sequence-359391-        6522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADH1t:HIS3_23∆29-XII-5 :.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6522)
  AUTHORS   Jensen ED, Laloux M, Lehka BJ, Pedersen LE, Jakociunas T, Jensen MK,
            Keasling JD
  TITLE     A synthetic RNA-mediated evolution system in yeast.
  JOURNAL   Nucleic Acids Res. 2021 Sep 7;49(15):e88. doi: 10.1093/nar/gkab472.
  PUBMED    34107026
REFERENCE   2  (bases 1 to 6522)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2021-09-7"; volume: "49(15):e88. doi"; pages: " 
            10.1093/nar/gkab472"
COMMENT     Sequence Label: pEDJ506
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..29)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11..29)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(39..55)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(48..70)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(834..867)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1337..1353)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1452..1469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1623..2211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1703..1722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2333..3124,3125..3193))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2956..2975
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3194..3285)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3327..3346
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3352..3371
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    3578..3611
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     terminator      complement(4144..4309)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     terminator      4459..4624
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     terminator      6326..6515
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
ORIGIN
        1 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
       61 cgtgactggg aaaaccctgg cgttacccct gcaggactag tgctgaggca ttaatgcaac
      121 tcagaagttt gacagcaagc aagttcatca ttcgaactag ccttattgtt ttagttcagt
      181 gacagcgaac tgccgtactc gatgctttat ttctcacggt agagcggaag aacagatagg
      241 ggcagcgtga gaagagttag aaagtaaatt tttatcacgt ctgaagtatt cttattcata
      301 ggaaattttg caaggttttt tagctcaata acgggctaag ttatataagg tgttcacgcg
      361 attttcttgt tatgtatacc tcttctctga ggaatggtac tactgtcctg atgtaggctc
      421 cttaaattgg tgggcaagaa taacttatcg atattttgta tattggtctt ggagttcacc
      481 acgtaatgcc tgtttaagac catcagttaa ctctagtatt atttggtctt ggctactggc
      541 cgtttgctat tattcaagtc ttttgtgcct tcccgtcggg taagggagtt atttagggat
      601 acagaatcta acgaaaacta aatctcaatg attaactcta tttaatcctt ttttgaaagg
      661 caaaagaggt cccttgttca cttacaacgt tcttagccaa attcgcttat cacttactac
      721 ttcacgatat acagaagtaa aaacatataa aaagatgtct gtttgtttag ccatcacaaa
      781 aggtatcgca gattaagacc tcagcgcggc cgcaaattta aataaaatga agtgaagttc
      841 ctatactttc tagagaatag gaacttctat agtgagtcga ataagggcga cacaaaattt
      901 attctaaatg cataataaat actgataaca tcttatagtt tgtattatat tttgtattat
      961 cgttgacatg tataattttg atatcaaaaa ctgattttcc ctttattatt ttcgagattt
     1021 attttcttaa ttctctttaa caaactagaa atattgtata tacaaaaaat cataaataat
     1081 agatgaatag tttaattata ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag
     1141 tgcgttgctt ttttctcatt tataaggtta aataattctc atatatcaag caaagtgaca
     1201 ggcgccctta aatattctga caaatgctct ttccctaaac tccccccata aaaaaacccg
     1261 ccgaagcggg tttttacgtt atttgcggat taacgattac tcgttatcag aaccgcccag
     1321 ggggcccgag cttaagactg gccgtcgttt tacaacacag aaagagtttg tagaaacgca
     1381 aaaaggccat ccgtcagggg ccttctgctt agtttgatgc ctggcagttc cctactctcg
     1441 ccttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2041 gctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2221 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag
     2281 ggattttggt catgagcttg cgccgtcccg tcaagtcagc gtaatgctct gcttaccaat
     2341 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2401 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagcgctg
     2461 cgatgatacc gcgagaacca cgctcaccgg ctccggattt atcagcaata aaccagccag
     2521 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2581 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2641 ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2701 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2761 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2821 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2881 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2941 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3001 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3061 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3121 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3181 gttgaatact catattcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3241 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gtcagtgtta
     3301 caaccaatta accaattctg aacattatcg cgagcccatt tatacctgaa tatggctcat
     3361 aacacccctt gtttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     3421 tcagaagtga aacgccgtag cgccgatggt agtgtgggga ctccccatgc gagagtaggg
     3481 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgcccggg
     3541 ctaattatgg ggtgtcgccc ttattcgact ctatagtgaa gttcctattc tctagaaagt
     3601 ataggaactt ctgaagtggg gatttaaatg cggccgcgct gagggtttaa tcaatctggc
     3661 ggcttgagtt ctcaacatgt tttatttttt acttatattg ctggtagggt aaaaaaatat
     3721 aactcctagg aataggttgt ctatatgttt ttgtcttgct tctataattg taacaaacaa
     3781 ggaaagggaa aatactgggt gtaaaagcca ttgagtcaag ttaggtcatc ccttttatac
     3841 aaaatttttc aatttttttt ccaagattct tgtacgatta attatttttt ttttgcgtcc
     3901 tacagcgtga tgaaaatttc gcctgctgca agatgagcgg gaacgggcga aatgtgcacg
     3961 cgcacaactt acgaaacgcg gatgagtcac tgacagccac cgcagaggtt ctgactccta
     4021 ctgagctcta ttggaggtgg cagaaccggt accggaggag gccgctataa ccggtttgaa
     4081 tttattgtca cagtgtcaca tcagcattaa gtcctcagcg agctcgcatg gaatgcgtgc
     4141 gatgagcgac ctcatgctat acctgagaaa gcaacctgac ctacaggaaa gagttactca
     4201 agaataagaa ttttcgtttt aaaacctaag agtcacttta aaatttgtat acacttattt
     4261 tttttataac ttatttaata ataaaaatca taaatcataa gaaattcgct tatttagaag
     4321 tgtcaacaac gtatctacca acggaatgcg tgcgatcgtt ggtagatacg ttgttgacac
     4381 ttctaaataa gcgaatttcc ttcgatcaac tcgcgccgct ggtccgttgg tagatacgtt
     4441 gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
     4501 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     4561 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc
     4621 gctcgttaat taagagctcg aatgcatctg tcatctacat aagaacacct ttggtggagg
     4681 gaacatcgtt ggtaccattg ggcgaggtgg cttctcttat ggcaaccgca agagccttga
     4741 acgcactctc actacggtga tgatcattct tgcctcgcag acaatcaacg tggagggtaa
     4801 ttctgctagc ctctgcaaag ctttcaagaa aatgcgggat catctcgcaa gagagatctc
     4861 ctactttctc cctttgcaaa ccaagttcga caactgcgta cggcctgttc gaaagatcta
     4921 caaccgctct ggaaagtgcc tcatccaaag gcgcaaatcc tgatccaaac ctttttactc
     4981 cacgcacggc ccctagggcc tctttaaaag cttgaccgag agcaatcccg cagtcttcag
     5041 tggtgtgatg gtcgtctatg tgtaagtcac caatgcactc ccgtcgagga atttttacta
     5101 acaccagagc atgtatcata tggtccagaa accctatacc tgtgtggacg ttaatcactt
     5161 gcgattgtgt ggcctgttct gctactgctt ctgcctcttt ttctgggaag atcgagtgct
     5221 ctatcgctag gggaccaccc tttaaagaga tcgcaatctg aatcttggtt tcatttgtaa
     5281 tacgctttac tagggctttc tgctctgtca ttgttttagt gcaggttgtt ttatatttgt
     5341 tgtaaaaagt agataattac ttccttgatg atctgtaaaa aagagaaaaa gaaagcatct
     5401 aagaacttga aaaactacga attagaaaag accaaatatg tatttcttgc attgaccaat
     5461 ttatgcaagt ttatatatat gtaaatgtaa gtttcacgag gttctactaa actaaaccac
     5521 ccccttggtt agaagaaaag agtgtgtgag aacaggctgt tgttgtcaca cgattcggac
     5581 aattctgttt gaaagagaga gagtaacagt acgatcgaac gaactttgct ctggagatca
     5641 cagtgggcat catagcatgt ggtactaaac cctttcccgc cattccagaa ccttcgattg
     5701 cttgttacaa aacctgtgag ccgtcgctag gaccttgttg tgtgacgaaa ttggaagctg
     5761 caatcaatag gaagacagga agtcgagcgt gtctgggttt tttcagtttt gttctttttg
     5821 caaacaaatc acgagcgacg gtaatttctt tctcgataag aggccacgtg ctttatgagg
     5881 gtaacatcaa ttcaagaagg agggaaacac ttcctttttc tggccctgat aatagtatga
     5941 gggtgaagcc aaaataaagg attcgcgccc aaatcggcat ctttaaatgc aggtatgcga
     6001 tagttcctca ctctttcctt actcacgagt aattcttgca aatgcctatt atgcagatgt
     6061 tataatatct gtgcgtcttg agttgaagtc aggaatctaa aataaaaatt aaggttaata
     6121 aaaagaggaa agaaaaaaaa attaatcgat ttacagaaac ttgcacacta aaaatacaca
     6181 actaaaagca attacagtat gggaagtcat cgacgttatc tctactatag tatattatca
     6241 tttctattat tatcctgctc agtggtactt gcaaaacaag ataagacccc attctttgaa
     6301 ggtacttctt cgatcgcgtg cattcatccg ctctaaccga aaaggaagga gttagacaac
     6361 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     6421 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     6481 aaaccttgct tgagaaggtt ttgggacgct cgaagatcgc gt
//
