LOCUS       sequence-97760-e        2908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Plac driving expression of GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2908)
  AUTHORS   Shis DL, Hussain F, Meinhardt S, Swint-Kruse L, Bennett MR
  TITLE     Modular, Multi-Input Transcriptional Logic Gating with Orthogonal 
            LacI/GalR Family Chimeras.
  JOURNAL   ACS Synth Biol. 2014 Sep 19;3(9):645-51. doi: 10.1021/sb500262f. 
            Epub 2014 Jul 28.
  PUBMED    25035932
REFERENCE   2  (bases 1 to 2908)
  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"; doi: "10.1021/sb500262f";
            journalName: "ACS Synth Biol"; date: "2014-09-19- 19"; volume: "3"; 
            issue: "9"; pages: "645-51"
COMMENT     Sequence Label: pZA2[PLac:GFP]
FEATURES             Location/Qualifiers
     source          1..2908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    9..25
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    32..48
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             108..824
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      863..949
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(985..1002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1119..1664)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1199..1218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1778..1872)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1903..2697)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2014..2033)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2624..2643
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2876..2894
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaatt gtgagcggat aacaattgac attgtgagcg gataacaaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccatg tctaaaggtg
      121 aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc
      181 acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa
      241 aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc actactttaa
      301 cttatggtgt tcaatgtttt tctagatacc cagatcatat gaaacaacat gactttttca
      361 agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta
      421 actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat
      481 taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact
      541 ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc aaagttaact
      601 tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat tatcaacaaa
      661 atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta tccactcaat
      721 ctaaattatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttgtta
      781 ctgctgctgg tattacccat ggtatggatg aattgtacaa atgaaagctt aattagctga
      841 tctagacgcg tgctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
      901 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
      961 gacctagggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     1021 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     1081 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
     1141 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     1201 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     1261 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     1321 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     1381 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     1441 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     1501 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     1561 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     1621 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     1681 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     1741 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     1801 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     1861 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
     1921 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     1981 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     2041 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     2101 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     2161 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     2221 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     2281 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     2341 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     2401 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     2461 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     2521 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     2581 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     2641 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     2701 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     2761 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     2821 agctggcaat tccgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2881 gcgtatcacg aggccctttc gtcttcac
//
