LOCUS       sequence-99929-e        8114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAPT110 encoding rbsH-hrpR-ter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8114)
  AUTHORS   Wang B, Kitney RI, Joly N, Buck M
  TITLE     Engineering modular and orthogonal genetic logic gates for robust 
            digital-like synthetic biology.
  JOURNAL   Nat Commun. 2011 Oct 18;2:508. doi: 10.1038/ncomms1516.
  PUBMED    22009040
REFERENCE   2  (bases 1 to 8114)
  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: "Nat 
            Commun."; date: "2011-10-18"; volume: "2:508. doi"; pages: " 
            10.1038/ncomms1516"
COMMENT     Sequence Label: pBW115lac-hrpR
FEATURES             Location/Qualifiers
     source          1..8114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    48..69
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(84..89,90..107,108..114)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    122..138
                     /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)."
     primer_bind     127..149
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1072..1083
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      1124..1195
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1211..1238
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1245..1265
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             2155..2946
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        3346..3423
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3424..4506
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3443..3462)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      4996..5541
                     /direction=RIGHT
                     /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     5442..5461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5658..5675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6125..6144)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6136..6951
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             6136..6951
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6209..6228)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      7395..7481
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7576..7662
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7757..7843
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7938..8024
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaattcccgg ggatctcgag gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
       61 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      121 attgtgagcg gataacaatt tcacacaggt ctagaaggag atataccatg agtacaggca
      181 tcgataagga cgtccgagag tgttggggcg taactgcatt atcagcgggt catcaaattg
      241 caatgaatag cgcgtttctg gatatggact tgctgttgtg cggggaaacc ggcaccggca
      301 aggacacact ggccaaccgc attcacgagt tgtccagcag gtcgggaccc tttgtgggca
      361 tgaactgcgc cgccattccc gagtcgctgg cagagagcca gttattcggt gtggtcaacg
      421 gtgcattcac cggcgtatgc cgggctcgcg agggctacat agaggcctcc agtggtggca
      481 ccttgtacct ggatgaaatc gacagcatgc cgttgagcct gcaagccaaa ctgctgcgtg
      541 tgttggagag tcgaggtatc gagcgtctgg gctcgaccga atttatcccg gtggatctgc
      601 ggatcattgc ctcggcccag cggccactgg atgaactggt ggaacaagga cttttccgtc
      661 gcgacctgtt ttttcggctc aacgtgctga cgcttcactt gccagccttg cgcaaacgtc
      721 gtgaacagat cctgccattg ttcgaccagt tcacccaggg tatcgctgcc gagttcggac
      781 gtcccgctcc tgcgctggac agcgggcgtg tgcagctgct gctcagccac gactggccgg
      841 gcaacatccg cgaattgaag tctgcggcca agcgcttcgt actcggcttc cccttgctgg
      901 gcgccgaccc tgtggaagcg cttgaccctg ccacggggct gcgcacgcaa atgcgcatca
      961 tcgagaaaat gctcatccag gatgccttga agcggcacag gcacaatttc gacgcggtgc
     1021 ttcaggagtt ggagttgcca agacgcaccc tgtatcaccg catgaaggaa ctgggagttg
     1081 cagcgccgat cgctgcgacg gccggggtct aataaccagg catcaaataa aacgaaaggc
     1141 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     1201 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag tagcggccgc
     1261 tgcagggtac cgagctcgaa ttgatccggt gattgattga cgaagcttta tgcttgtaaa
     1321 ccgttttgtg aaaaaatttt taaaataaaa aaggggacct ctagggtccc caattaatta
     1381 gtaatataat ctattaaagg tcattcaaaa ggtcatccac cggatcaatt cccctgctcg
     1441 cgcaggctgg gtgccaagct ctcgggtaac atcaaggccc gatccttgga gcccttgccc
     1501 tcccgcacga tgatcgtgcc gtgatcgaaa tccagatcct tgacccgcag ttgcaaaccc
     1561 tcactgatcc gcatgcccgt tccatacaga agctgggcga acaaacgatg ctcgccttcc
     1621 agaaaaccga ggatgcgaac cacttcatcc ggggtcagca ccaccggcaa gcgccgcgac
     1681 ggccgaggtc ttccgatctc ctgaagccag ggcagatccg tgcacagcac cttgccgtag
     1741 aagaacagca aggccgccaa tgcctgacga tgcgtggaga ccgaaacctt gcgctcgttc
     1801 gccagccagg acagaaatgc ctcgacttcg ctgctgccca aggttgccgg gtgacgcaca
     1861 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
     1921 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     1981 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
     2041 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     2101 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg
     2161 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     2221 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     2281 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     2341 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     2401 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     2461 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     2521 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     2581 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     2641 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     2701 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     2761 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     2821 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     2881 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     2941 aaataatgtc taacaattcg ttcaagccga cgccgcttcg cggcgcggct taactcaagc
     3001 gttagatgca ctaagcacat aattgctcac agccaaacta tcaggtcaag tctgctttta
     3061 ttatttttaa gcgtgcataa taagccctac acaaattggg agatatatca tgaaaggctg
     3121 gctttttctt gttatcgcaa tagttggcga agtaatcgca acatccgcat taaaatctag
     3181 cgagggcttt actaagctga tccggtggat gaccttttga atgaccttta atagattata
     3241 ttactaatta attggggacc tagaggtccc cttttttatt ttaaaaattt tttacaaaac
     3301 ggtttacaag cataaagctt gctcaatcaa tcaccggatc aattcgacac catcgaatgg
     3361 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     3421 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     3481 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     3541 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     3601 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     3661 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     3721 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     3781 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     3841 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     3901 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     3961 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     4021 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     4081 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     4141 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     4201 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     4261 tcatgttata tcccgccgtc aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     4321 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     4381 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     4441 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4501 cagtgagcga cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4681 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4801 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4861 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4921 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagacccctt
     4981 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
     5041 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg
     5101 aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc
     5161 ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg
     5221 cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5281 cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac
     5341 tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac
     5401 cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt
     5461 tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca
     5521 ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta
     5581 tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca
     5641 gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca
     5701 tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac
     5761 accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg aggtctgcct
     5821 cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa
     5881 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac
     5941 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
     6001 tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac
     6061 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     6121 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta
     6181 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     6241 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     6301 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     6361 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     6421 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     6481 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     6541 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     6601 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     6661 gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact
     6721 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     6781 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     6841 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     6901 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     6961 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt
     7021 gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag
     7081 accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc
     7141 tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg
     7201 agtcagcaac accttcttca cgaggcagac ctcagcgcta ttctgacctt gccatcacga
     7261 ctgtgctggt cattaaacgc gtattcaggc tgaccctgcg cgctgcgcag ggctttattg
     7321 attccatttt tacactgatg aatgttccgt tgcgctgccc ggattacagc ggatcaattc
     7381 ccaattccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     7441 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     7501 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga
     7561 attaattcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     7621 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     7681 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     7741 aattaattcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     7801 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     7861 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     7921 gaattaattc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     7981 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     8041 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     8101 ggaattgggg atcg
//
