LOCUS       sequence-9606-ex        5747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5747)
  AUTHORS   Hackett EA, Esch RK, Maleri S, Errede B
  TITLE     A family of destabilized cyan fluorescent proteins as 
            transcriptional reporters in S. cerevisiae.
  JOURNAL   Yeast. 2006 Apr 15. 23(5):333-49.
  PUBMED    16598699
REFERENCE   2  (bases 1 to 5747)
  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: "Yeast. 2006
            Apr 15. 23(5):333-49."
COMMENT     Sequence Label: pNC891
FEATURES             Location/Qualifiers
     source          1..5747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..463
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             494..1207
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(521..549)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1144..1164
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1228..1415
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1283..1307)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1490..2846
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1490..1833
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1834..2643
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1901..1920)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2649..2846
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3384..3403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3385..3403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3490..3507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3661..4249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3741..3760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4420..5211,5212..5280))
                     /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     5043..5062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5281..5385)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5453..5471
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5509..5531)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5631..5650
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5731..5747,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5731..5747,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg actattgaag taataatagg
       61 cgcatgcaac ttcttttctt tttttttctt ttctctctcc cccgttgttg tctcaccata
      121 tccgcaatga caaaaaaatg atggaagaca ctaaaggaaa aaattaacga caaagacagc
      181 accaacagat gtcgttgttc cagagctgat gaggggtatc tcgaagcaca cgaaactttt
      241 tccttccttc attcacgcac actactctct aatgagcaac ggtatacggc cttccttcca
      301 gttacttgaa tttgaaataa aaaaaagttt gctgtcttgc tatcaagtat aaatagacct
      361 gcaattatta atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt
      421 tttctgcaca atatttcaag ctataccaag catacaatca actatctcat atacaatgcc
      481 cgggttaatt aacagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
      541 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
      601 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
      661 gccaacactt gtcactactt tcggttatgg tctaaaatgc tttgctagat acccagatca
      721 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
      781 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
      841 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
      901 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
      961 aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca
     1021 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     1081 caaccattac ctgtcctatc aatctgccct ttcgaaagat cccaacgaaa agagagacca
     1141 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     1201 caaatagggc gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt
     1261 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     1321 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     1381 tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga taacagggta
     1441 atatagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc
     1501 ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg
     1561 cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg
     1621 ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc
     1681 tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg
     1741 ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata
     1801 cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg
     1861 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1921 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     1981 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2041 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2101 gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa
     2161 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2221 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2281 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2341 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     2401 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2461 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2521 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2581 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2641 taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt
     2701 tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc
     2761 tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc
     2821 gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt
     2881 aaacgagctc aaaaaggtgt tatcttctac gaatcccacg gtaagttgga atacaaagat
     2941 attccagttc caaagccaaa ggccaacgaa ttgttgatca acgttaaata ctctggtgtc
     3001 tgtcacactg acttgcacgc ttggcacggt gactggccat tgccagttaa gctaccatta
     3061 gtcggtggtc acgaaggtgc cggtgtcgtt gtcggcatgg gtgaaaacgt taagggctgg
     3121 aagatcggtg actacgccgg tatcaaatgg ttgaacggtt cttgtatggc ctgtgaatac
     3181 tgtgaattgg gtaacgaatc caactgtcct cacgctgact tgtctggtta cacccacgac
     3241 ggttctttcc aacaatacgc taccgctgac gctgttcaag ccgctcacat tcctcaaggt
     3301 accgacttgg cccaagtcgc ccccatcttg tgtgctgata tcagatccac tagtggccta
     3361 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     3421 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3841 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5401 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5461 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5521 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5581 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5641 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
     5701 atacataacc ttatgtatca tacacatacg atttaggtga cactata
//
