LOCUS       sequence-419161-        7987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for biosynthetic pathway. pTac-phiC31 integrase. This 
            material contains 2 plasmids. For more information look under the 
            ""Depositor Comments"" section below..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 6652 to 6686)
  AUTHORS   Patel JR, Oh J, Wang S, Crawford JM, Isaacs FJ
  TITLE     Cross-kingdom expression of synthetic genetic elements promotes 
            discovery of metabolites in the human microbiome.
  JOURNAL   Cell. 2022 Apr 28;185(9):1487-1505.e14. doi: 
            10.1016/j.cell.2022.03.008. Epub 2022 Apr 1.
  PUBMED    35366417
REFERENCE   2  (bases 1 to 7987)
  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: 
            "10.1016/j.cell.2022.03.008"; journalName: "Cell"; date: 
            "2022-04-28- 28"; volume: "185"; issue: "9"; pages: "1487-1505.e14"
COMMENT     Sequence Label: pPath-pTac-integrase
FEATURES             Location/Qualifiers
     source          1..7987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..37
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        18..36
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             76..531
                     /codon_start=1
                     /product="lysozyme from bacteriophage T7"
                     /label=T7 lysozyme
                     /note="inhibitor of T7 RNA polymerase"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAVLRAHHEVAPKACPSFDLKRWWEKNELVTSDRG"
     terminator      560..646
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(681..698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(815..1360)
                     /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(895..914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1525..2316)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFGPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        2786..2863
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2864..3946
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2883..2902)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    3959..3980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3995..4000,4001..4018,4019..4025)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4033..4049
                     /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     4038..4060
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        6652..6686
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
ORIGIN
        1 aagcttgatg ggggatctaa tacgactcac tatagggaga tttgatagat taaaaaggaa
       61 aggaggaaag aaataatggc tcgtgtacag tttaaacaac gtgaatctac tgacgcaatc
      121 tttgttcact gctcggctac caagccaagt cagaatgttg gtgtccgtga gattcgccag
      181 tggcacaaag agcagggttg gctcgatgtg ggataccact ttatcatcaa gcgagacggt
      241 actgtggagg caggacgaga tgagatggct gtaggctctc acgctaaggg ttacaaccac
      301 aactctatcg gcgtctgcct tgttggtggt atcgacgata aaggtaagtt cgacgctaac
      361 tttacgccag cccaaatgca atcccttcgc tcactgcttg tcacactgct ggctaagtac
      421 gaaggcgctg tgcttcgcgc ccatcatgag gtggcgccga aggcttgccc ttcgttcgac
      481 cttaagcgtt ggtgggagaa gaacgaactg gtcacttctg accgtggata agatcccatg
      541 gtacgcgtgc tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      601 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac
      661 ctaggggtat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
      721 tgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
      781 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggcttcgc ccccctgaca
      841 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
      901 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
      961 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagtccc
     1021 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     1081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     1141 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     1201 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     1261 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     1321 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     1381 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     1441 ccccatacga tataagttgt tactagttgc agaaataaaa aggcctgcga ttaccggcag
     1501 tcctgttatt agctcagtaa agctttattt gccgactacc ttggtgatct cgcctttcac
     1561 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     1621 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     1681 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     1741 gcgggacaac gtaagcacta catttcgctc atcgcctgcc cagtcgggcg gcgagttcca
     1801 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     1861 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     1921 agccagatca atgtcgatcg tggctggccc gaagatacct gcaagaatgt cattgcgctg
     1981 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     2041 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     2101 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     2161 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     2221 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     2281 ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg
     2341 actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     2401 acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca
     2461 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     2521 gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca
     2581 ctgggttcgt gccttcatcc gtttccacgg tgtgtgtcac ccggcaacct tgggtagcag
     2641 cgaagtcgag gcatttctgt cctggctgag atcttgatcc cctgcgccat cagatccttg
     2701 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgcga
     2761 aggcgaagcg gcatgcattt acgttgacac catcgaatgg cgcaaaacct ttcgcggtat
     2821 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     2881 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     2941 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     3001 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     3061 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     3121 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     3181 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     3241 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     3301 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     3361 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     3421 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     3481 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     3541 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     3601 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     3661 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     3721 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     3781 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     3841 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3901 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3961 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     4021 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gtttaatggt
     4081 ggcattcgtc gcttcgaagc agatcaacaa cgccagattg cagcaggtag cgagagcgac
     4141 acagcatgga accgccgcct gttgtcagaa cttattgcac ctatggctga aggcattcag
     4201 gcttataaag aagagtacga aggtaagaaa ggtcgtgcac ctcgcgcatt ggctttctta
     4261 caatgtgtag aaaatgaagt tgcagcatac atcactatga aagttgttat ggatatgctg
     4321 aatacggatg ctacccttca ggctattgca atgagtgtag cagaacgcat tgaagaccaa
     4381 gtgcgctttt ctaagctaga aggtcacgcc gctaaatact ttgagaaggt taagaagtca
     4441 ctcaaggcta gccgtactaa gtcatatcgt cacgctcata acgtagctgt agttgctgaa
     4501 aaatcagttg cagaaaagga cgcggacttt gaccgttggg aggcgtggcc aaaagaaact
     4561 caattgcaga ttggtactac cttgcttgaa atcttagaag gtagcgtttt ctataatggt
     4621 gaacctgtat ttatgcgtgc tatgcgcact tatggcggaa agactattta ctacttacaa
     4681 acttctgaaa gtgtaggcca gtggattagc gcattcaaag agcacgtagc gcaattaagc
     4741 ccagcttatg ccccttgcgt aatccctcct cgtccttgga gaactccatt taatggaggg
     4801 ttccatactg agaaggtagc tagccgtatc cgtcttgtaa aaggtaaccg tgagcatgta
     4861 cgcaagttga ctcaaaagca aatgccaaag gtttataagg ctatcaacgc attacaaaat
     4921 acacaatggc aaatcaacaa ggatgtatta gcagttattg aagaagcaat ccgcttagac
     4981 cttggttatg gtgtaccttc cttcaagcca ctgattgaca aggagaacaa gccagctaac
     5041 ccggtacctg ttgaattcca acacctgcgc ggtcgtgaac tgaaagagat gctatcacct
     5101 gagcagtggc aacaattcat taactggaaa ggcgaatgcg cgcgcctata taccgcagaa
     5161 actaagcgcg gttcaaagtc cgccgccgtt gttcgcatgg taggacaggc ccgtaaatat
     5221 agcgcctttg aatccattta cttcgtgtac gcaatggata gccgcagccg tgtctatgtg
     5281 caatctagca cgctctctcc gcagtctaac gacttaggta aggcattact ccgctttacc
     5341 gagggacgcc ctgtgaatgg cgtagaagcg cttaaatggt tctgcatcaa tggtgctaac
     5401 ctttggggat gggacaagaa aacttttgat gtgcgcgtgt ctaacgtatt agatgaggaa
     5461 ttccaagata tgtgtcgaga catcgccgca gaccctctca cattcaccca atgggctaaa
     5521 gctgatgcac cttatgaatt cctcgcttgg tgctttgagt atgctcaata ccttgatttg
     5581 gtggatgaag gaagggccga cgaattccgc actcacctac cagtacatca ggacgggtct
     5641 tgttcaggca ttcagcacta tagtgctatg cttcgcgacg aagtaggggc caaagctgtt
     5701 aacctgaaac cctccgatgc accgcaggat atctatgggg cggtggcgca agtggttatc
     5761 aagaagaatg cgctatatat ggatgcggac gatgcaacca cgtttacttc tggtagcgtc
     5821 acgctgtccg gtacagaact gcgagcaatg gctagcgcat gggatagtat tggtattacc
     5881 cgtagcttaa ccaaaaagcc cgtgatgacc ttgccatatg gttctactcg cttaacttgc
     5941 cgtgaatctg tgattgatta catcgtagac ttagaggaaa aagaggcgca gaaggcagta
     6001 gcagaagggc ggacggcaaa caaggtacat ccttttgaag acgatcgtca agattacttg
     6061 actccgggcg cagcttacaa ctacatgacg gcactaatct ggccttctat ttctgaagta
     6121 gttaaggcac cgatagtagc tatgaagatg atacgccagc ttgcacgctt tgcagcgaaa
     6181 cgtaatgaag gcctgatgta caccctgcct actggcttca tcttagaaca gaagatcatg
     6241 gcaaccgaga tgctacgcgt gcgtacctgt ctgatgggtg atatcaagat gtcccttcag
     6301 gttgaaacgg atatcgtaga tgaagccgct atgatgggag cagcagcacc taatttcgta
     6361 cacggtcatg acgcaagtca ccttatcctt accgtatgtg aattggtaga caagggcgta
     6421 actagtatcg ctgtaatcca cgactctttt ggtactcatg cagacaacac cctcactctt
     6481 agagtggcac ttaaagggca gatggttgca atgtatattg atggtaatgc gcttcagaaa
     6541 ctactggagg agcatgaaga gcgctggatg gttgatacag gtatcgaagt acctgagcaa
     6601 ggggagttcg accttaacga aatcatggat tctgaatacg tatttgccta attgacggct
     6661 agctcagtcc taggtacagt gctagcagct aaagctatat aatttaatta ggagaagtaa
     6721 aatgcaggaa ggcgcgtatc gttttattcg taatccgaac gtgagcgcgg aagcgattcg
     6781 taaagcgggt gccatgcaga ccgtgaaact ggcccaggaa tttccggaac tgctggcaat
     6841 tgaagatacc acctctctga gctatcgtca tcaggtggcg gaagaactgg gcaaactggg
     6901 tagcattcag gataaaagcc gtggttggtg ggtgcatagc gtgctgctgc tggaagcgac
     6961 cacctttcgt accgtgggcc tgctgcatca agaatggtgg atgcgtccgg atgatccggc
     7021 ggatgcggat gaaaaagaaa gcggcaaatg gctggccgct gctgcaactt cgcgtctgag
     7081 aatgggcagc atgatgagca acgtgattgc ggtgtgcgat cgtgaagcgg atattcatgc
     7141 gtatctgcaa gataaactgg cccataacga acgttttgtg gtgcgtagca aacatccgcg
     7201 taaagatgtg gaaagcggcc tgtatctgta tgatcacctg aaaaaccagc cggaactggg
     7261 cggctatcag attagcattc cgcagaaagg cgtggtggat aaacgtggca aacgtaaaaa
     7321 ccgtccggcg cgtaaagcga gcctgagcct gcgtagcggc cgtattaccc tgaaacaggg
     7381 caacattacc ctgaacgcgg tgctggccga agaaattaat ccgccgaaag gcgaaacccc
     7441 gctgaaatgg ctgctgctga ccagcgagcc ggtggaaagt ctggcccaag cgctgcgtgt
     7501 gattgatatt tatacccatc gttggcgcat tgaagaattt cacaaagcgt ggaaaacggg
     7561 tgcgggtgcg gaacgtcagc gtatggaaga accggataac ctggaacgta tggtgagcat
     7621 tctgagcttt gtggcggtgc gtctgctgca actgcgtgaa tcttttactc cgccgcaagc
     7681 actgcgtgcg cagggcctgc tgaaagaagc ggaacacgtt gaaagccaga gcgcggaaac
     7741 cgtgctgacc ccggatgaat gccaactgct gggctatctg gataaaggca aacgcaaacg
     7801 caaagaaaaa gcgggcagcc tgcaatgggc gtatatggcg attgcgcgtc tgggcggctt
     7861 tatggatagc aaacgtaccg gcattgcgag ctggggtgcg ctgtgggaag gttgggaagc
     7921 gctgcaaagc aaactggatg gctttctggc cgcgaaagac ctgatggcgc agggcattaa
     7981 aatctaa
//
