LOCUS       sequence-60893-e        5941 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 5941)
  AUTHORS   Nevozhay D, Adams RM, Van Itallie E, Bennett MR, Balazsi G
  TITLE     Mapping the environmental fitness landscape of a synthetic gene 
            circuit.
  JOURNAL   PLoS Comput Biol. 2012;8(4):e1002480. doi: 
            10.1371/journal.pcbi.1002480. Epub 2012 Apr 12.
  PUBMED    22511863
REFERENCE   2  (bases 1 to 5941)
  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: "PLoS Comput
            Biol."; date: "2012"; volume: "8(4):e1002480. doi"; pages: " 
            10.1371/journal.pcbi.1002480. Epub 2012 Apr 12"
COMMENT     Sequence Label: pDN-T2dGZmxh
FEATURES             Location/Qualifiers
     source          1..5941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        383..570
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             571..1233
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1493..1948)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1630..1651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1842..1861
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      2050..2237
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2105..2129)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2282..2300
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2324..2342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2545..3258
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3283..3657
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3690..3879
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3951..3968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4122..4710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4202..4221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4881..5672,5673..5741))
                     /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     5504..5523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5742..5846)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5914..5932
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcat tgtttttttc tactattgct ttgctgtggg aaaaacttat cgaaagatga
      241 cgactttttc ttaattctcg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta
      301 tcgtttgaac acggcattag tcagggaagt cataacacag tcctttcccg caattttctt
      361 tttctattac tcttggcctc ctctagtaca ctctatattt ttttatgcct cggtaatgat
      421 tttcattttt ttttttccac ctagcggatg actctttttt tttcttagcg attggcatta
      481 tcacataatg aattatacat tatataaagt aatgtgattt cttcgaagaa tatactaaaa
      541 aatgagcagg caagataaac gaaggcaaag atgacagagc agaaagccct agtaaagcgt
      601 attacaaatg aaaccaagat tcagattgcg atctctttaa agggtggtcc cctagcgata
      661 gagcactcga tcttcccaga aaaagaggca gaagcagtag cagaacaggc cacacaatcg
      721 caagtgatta acgtccacac aggtataggg tttctggacc atatgataca tgctctggcc
      781 aagcattccg gctggtcgct aatcgttgag tgcattggtg acttacacat agacgaccat
      841 cacaccactg aagactgcgg gattgctctc ggtcaagctt ttaaagaggc cctaggggcc
      901 gtgcgtggag taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga
      961 gcggtggtag atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag
     1021 aaagtaggag atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct
     1081 agcagaatta ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag
     1141 agtgcgttca aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac
     1201 gatgttccct ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca
     1261 tacgatatat atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga
     1321 acagtatgat actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga
     1381 ggcgcgcttt ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc
     1441 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     1501 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     1561 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     1621 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     1681 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     1741 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     1801 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     1861 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     1921 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     1981 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggagctctta aggcgccact
     2041 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     2101 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     2161 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt
     2221 gaccacacct ctaccggcag atccgctagg gataacaggg taatatagat caattcctcg
     2281 atccctatca gtgatagaga gtcgacaaag tcgagtttct cgatccctat cagtgataga
     2341 gagtcgacaa agtcgagttt ctcgatcgag accactgcat gcatgtgctc tgtatgtata
     2401 taaaactctt gttttcttct tttctctaaa tattctttcc ttatacatta ggtcctttgt
     2461 agcataaatt actatacttc tatagacacg caaacacaaa tacacacact aaattaccgg
     2521 atcaattcgg gggatcctat taaaatgtct aaaggtgaag aattattcac tggtgttgtc
     2581 ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa
     2641 ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa
     2701 ttgccagttc catggccaac cttagtcact actttcggtt atggtgttca atgttttgcg
     2761 agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat
     2821 gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc
     2881 aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa
     2941 gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc
     3001 atggctgaca aacaaaagaa tggtatcaaa gttaacttca aaattagaca caacattgaa
     3061 gatggttctg ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca
     3121 gtcttgttac cagacaacca ttacttatcc actcaatctg ccttatccaa agatccaaac
     3181 gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt
     3241 atggatgaat tgtacaaagg tgacggtgct ggtttaatta acatggccaa gttgaccagt
     3301 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     3361 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     3421 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     3481 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     3541 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     3601 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgactc
     3661 gagtaagctt ggtaccgcgg ctagctaaga tccgctctaa ccgaaaagga aggagttaga
     3721 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     3781 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     3841 ctgaaaacct tgcttgagaa ggttttggga cgctcgaaga tccagctgca ttaatgaatc
     3901 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4561 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4861 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4921 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4981 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5041 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5101 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5161 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5221 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5281 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5341 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5401 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5461 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5521 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5581 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5641 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5701 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5761 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5881 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5941 c
//
