LOCUS       sequence-97762-e        4957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PLac driving expression of mCherry and PTAN driving 
            expression of GFP. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4957)
  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 4957)
  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:mCherry,PTAN:GFP]
FEATURES             Location/Qualifiers
     source          1..4957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(452..479)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(664..1374)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(942..961)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1211..1229
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_RNA        complement(1410..1459)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    1502..1518
                     /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    1525..1541
                     /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             2139..2150
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2157..2873
                     /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      2912..2998
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3034..3051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3168..3713)
                     /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(3248..3267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3827..3921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3952..4746)
                     /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(4063..4082)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4673..4692
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4925..4943
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttgaaccatg gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
       61 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
      121 tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt
      181 ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg
      241 ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct
      301 tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg
      361 ctgcaccggt gcgcgatttc aacacgagcc aaaataataa aaaagccgga ttaataatct
      421 ggctttttat attctctctc tagtatataa acgcagaaag gcccacccga aggtgagcca
      481 gtgtgatcca agccagctcg atatccgcct tactggttag cagaatgaat caccgatacg
      541 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
      601 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtccccta cgtgctgccc
      661 gggtcacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
      721 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt
      781 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc
      841 gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc
      901 ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg
      961 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
     1021 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
     1081 ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt
     1141 cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag
     1201 ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     1261 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa
     1321 gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctaa ccattatcct
     1381 ttctccttat cggtaccttc tatatgaggc tgtttcgtcc tcacggactc atcagaccgg
     1441 aaagcacatc cggtgacagc tggcgcgccg gtcagtgcgt cctgctgatg tgctcagtat
     1501 cttgttatcc gctcacaatg tcaattgtta tccgctcaca attgcggccg cggcagttat
     1561 tggtgccctt aaacgcctgg tgctacgcct gaataagtga taataagcgg atgaatggca
     1621 gaaattcgaa agcaaattcg acccggtcgt cggttcaggg cagggtcgtt aaatagccgc
     1681 ttatgtctat tgctggttta ccggtttatt gactaccgga agcagtgtga ccgtgtgctt
     1741 ctcaaatgcc tgactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1801 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     1861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2041 cgttaagggc tcgagaattt taagcggctt aaaattgaca ttttaagcgc ttaaaatgat
     2101 actgagcaca tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccatgt
     2161 ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg
     2221 ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat
     2281 tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca accttagtca
     2341 ctactttaac ttatggtgtt caatgttttt ctagataccc agatcatatg aaacaacatg
     2401 actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag
     2461 atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata
     2521 gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg
     2581 aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca
     2641 aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt
     2701 atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat
     2761 ccactcaatc taaattatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag
     2821 aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa tgaaagctta
     2881 attagctgat ctagacgcgt gctagaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2941 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3001 gccgccctag acctagggga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     3061 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     3121 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     3181 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     3241 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     3301 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     3361 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     3421 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     3481 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     3541 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     3601 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     3661 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     3721 atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag
     3781 cacctgaagt cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa
     3841 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     3901 ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa
     3961 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     4021 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     4081 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     4141 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     4201 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     4261 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     4321 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     4381 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     4441 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     4501 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     4561 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     4621 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     4681 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     4741 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     4801 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     4861 gggcgcccca gctggcaatt ccgacgtcta agaaaccatt attatcatga cattaaccta
     4921 taaaaatagg cgtatcacga ggccctttcg tcttcac
//
