LOCUS       sequence-423477-        8764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Landing Pad with T7 RNAP circuit variant T15. pIP1443 seed promoter.
            pTac-himar transposase. In dap auxotrophic conjugation donor. This 
            material contains 2 plasmids. For more information look below..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 5496 to 5530)
  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 8764)
  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: pLP-pTac-himar-apraR
FEATURES             Location/Qualifiers
     source          1..8764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(369..1160)
                     /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        1630..1707
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1708..2790
                     /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(1727..1746)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    2803..2824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2839..2844,2845..2862,2863..2869)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2877..2893
                     /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     2882..2904
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        5496..5530
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     primer_bind     6849..6868
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6849..6867
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     mobile_element  6917..7684
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             7687..8139
                     /codon_start=1
                     /product="lysozyme from bacteriophage T7"
                     /label=T7 lysozyme
                     /note="inhibitor of T7 RNA polymerase"
                     /translation="ARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDVG
                     YHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSLR
                     SLLVTLLAKYEGAVLRAHHEVAPKACPSFDLKRWWEKNELVTSDRG"
     terminator      8168..8254
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(8289..8306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8423..8764,1..204))
                     /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(8503..8522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
       61 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
      121 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
      181 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
      241 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
      301 ttgttactag ttgcagaaat aaaaaggcct gcgattaccg gcagtcctgt tattagctca
      361 gtaaagcttt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
      421 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
      481 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
      541 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
      601 actacatttc gctcatcgcc tgcccagtcg ggcggcgagt tccatagcgt taaggtttca
      661 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
      721 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
      781 atcgtggctg gcccgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
      841 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
      901 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
      961 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     1021 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
     1081 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     1141 gcgatcaccg cttccctcat aatgtttaac tttgttttag ggcgactgcc ctgctgcgta
     1201 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
     1261 gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat gaaaaccgcc
     1321 actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata
     1381 cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt tcgtgccttc
     1441 atccgtttcc acggtgtgtg tcacccggca accttgggta gcagcgaagt cgaggcattt
     1501 ctgtcctggc tgagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     1561 ccagtttact ttgcagggct tcccaacctt accagagggc gcgaaggcga agcggcatgc
     1621 atttacgttg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
     1681 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     1741 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     1801 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     1861 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     1921 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     1981 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
     2041 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     2101 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     2161 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     2221 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     2281 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     2341 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     2401 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     2461 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     2521 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     2581 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     2641 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     2701 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2761 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2821 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2881 gagcggataa caatttcaca caggaaacag ctatgtttaa tggtggcatt cgtcgcttcg
     2941 aagcagatca acaacgccag attgcagcag gtagcgagag cgacacagca tggaaccgcc
     3001 gcctgttgtc agaacttatt gcacctatgg ctgaaggcat tcaggcttat aaagaagagt
     3061 acgaaggtaa gaaaggtcgt gcacctcgcg cattggcttt cttacaatgt gtagaaaatg
     3121 aagttgcagc atacatcact atgaaagttg ttatggatat gctgaatacg gatgctaccc
     3181 ttcaggctat tgcaatgagt gtagcagaac gcattgaaga ccaagtgcgc ttttctaagc
     3241 tagaaggtca cgccgctaaa tactttgaga aggttaagaa gtcactcaag gctagccgta
     3301 ctaagtcata tcgtcacgct cataacgtag ctgtagttgc tgaaaaatca gttgcagaaa
     3361 aggacgcgga ctttgaccgt tgggaggcgt ggccaaaaga aactcaattg cagattggta
     3421 ctaccttgct tgaaatctta gaaggtagcg ttttctataa tggtgaacct gtatttatgc
     3481 gtgctatgcg cacttatggc ggaaagacta tttactactt acaaacttct gaaagtgtag
     3541 gccagtggat tagcgcattc aaagagcacg tagcgcaatt aagcccagct tatgcccctt
     3601 gcgtaatccc tcctcgtcct tggagaactc catttaatgg agggttccat actgagaagg
     3661 tagctagccg tatccgtctt gtaaaaggta accgtgagca tgtacgcaag ttgactcaaa
     3721 agcaaatgcc aaaggtttat aaggctatca acgcattaca aaatacacaa tggcaaatca
     3781 acaaggatgt attagcagtt attgaagaag caatccgctt agaccttggt tatggtgtac
     3841 cttccttcaa gccactgatt gacaaggaga acaagccagc taacccggta cctgttgaat
     3901 tccaacacct gcgcggtcgt gaactgaaag agatgctatc acctgagcag tggcaacaat
     3961 tcattaactg gaaaggcgaa tgcgcgcgcc tatataccgc agaaactaag cgcggttcaa
     4021 agtccgccgc cgttgttcgc atggtaggac aggcccgtaa atatagcgcc tttgaatcca
     4081 tttacttcgt gtacgcaatg gatagccgca gccgtgtcta tgtgcaatct agcacgctct
     4141 ctccgcagtc taacgactta ggtaaggcat tactccgctt taccgaggga cgccctgtga
     4201 atggcgtaga agcgcttaaa tggttctgca tcaatggtgc taacctttgg ggatgggaca
     4261 agaaaacttt tgatgtgcgc gtgtctaacg tattagatga ggaattccaa gatatgtgtc
     4321 gagacatcgc cgcagaccct ctcacattca cccaatgggc taaagctgat gcaccttatg
     4381 aattcctcgc ttggtgcttt gagtatgctc aataccttga tttggtggat gaaggaaggg
     4441 ccgacgaatt ccgcactcac ctaccagtac atcaggacgg gtcttgttca ggcattcagc
     4501 actatagtgc tatgcttcgc gacgaagtag gggccaaagc tgttaacctg aaaccctccg
     4561 atgcaccgca ggatatctat ggggcggtgg cgcaagtggt tatcaagaag aatgcgctat
     4621 atatggatgc ggacgatgca accacgttta cttctggtag cgtcacgctg tccggtacag
     4681 aactgcgagc aatggctagc gcatgggata gtattggtat tacccgtagc ttaaccaaaa
     4741 agcccgtgat gaccttgcca tatggttcta ctcgcttaac ttgccgtgaa tctgtgattg
     4801 attacatcgt agacttagag gaaaaagagg cgcagaaggc agtagcagaa gggcggacgg
     4861 caaacaaggt acatcctttt gaagacgatc gtcaagatta cttgactccg ggcgcagctt
     4921 acaactacat gacggcacta atctggcctt ctatttctga agtagttaag gcaccgatag
     4981 tagctatgaa gatgatacgc cagcttgcac gctttgcagc gaaacgtaat gaaggcctga
     5041 tgtacaccct gcctactggc ttcatcttag aacagaagat catggcaacc gagatgctac
     5101 gcgtgcgtac ctgtctgatg ggtgatatca agatgtccct tcaggttgaa acggatatcg
     5161 tagatgaagc cgctatgatg ggagcagcag cacctaattt cgtacacggt catgacgcaa
     5221 gtcaccttat ccttaccgta tgtgaattgg tagacaaggg cgtaactagt atcgctgtaa
     5281 tccacgactc ttttggtact catgcagaca acaccctcac tcttagagtg gcacttaaag
     5341 ggcagatggt tgcaatgtat attgatggta atgcgcttca gaaactactg gaggagcatg
     5401 aagagcgctg gatggttgat acaggtatcg aagtacctga gcaaggggag ttcgacctta
     5461 acgaaatcat ggattctgaa tacgtatttg cctaattgac ggctagctca gtcctaggta
     5521 cagtgctagc agctaaagct atataattta attaggagaa gtaaaatgca ggaaggcgcg
     5581 tatcgtttta ttcgtaatcc gaacgtgagc gcggaagcga ttcgtaaagc gggtgccatg
     5641 cagaccgtga aactggccca ggaatttccg gaactgctgg caattgaaga taccacctct
     5701 ctgagctatc gtcatcaggt ggcggaagaa ctgggcaaac tgggtagcat tcaggataaa
     5761 agccgtggtt ggtgggtgca tagcgtgctg ctgctggaag cgaccacctt tcgtaccgtg
     5821 ggcctgctgc atcaagaatg gtggatgcgt ccggatgatc cggcggatgc ggatgaaaaa
     5881 gaaagcggca aatggctggc cgctgctgca acttcgcgtc tgagaatggg cagcatgatg
     5941 agcaacgtga ttgcggtgtg cgatcgtgaa gcggatattc atgcgtatct gcaagataaa
     6001 ctggcccata acgaacgttt tgtggtgcgt agcaaacatc cgcgtaaaga tgtggaaagc
     6061 ggcctgtatc tgtatgatca cctgaaaaac cagccggaac tgggcggcta tcagattagc
     6121 attccgcaga aaggcgtggt ggataaacgt ggcaaacgta aaaaccgtcc ggcgcgtaaa
     6181 gcgagcctga gcctgcgtag cggccgtatt accctgaaac agggcaacat taccctgaac
     6241 gcggtgctgg ccgaagaaat taatccgccg aaaggcgaaa ccccgctgaa atggctgctg
     6301 ctgaccagcg agccggtgga aagtctggcc caagcgctgc gtgtgattga tatttatacc
     6361 catcgttggc gcattgaaga atttcacaaa gcgtggaaaa cgggtgcggg tgcggaacgt
     6421 cagcgtatgg aagaaccgga taacctggaa cgtatggtga gcattctgag ctttgtggcg
     6481 gtgcgtctgc tgcaactgcg tgaatctttt actccgccgc aagcactgcg tgcgcagggc
     6541 ctgctgaaag aagcggaaca cgttgaaagc cagagcgcgg aaaccgtgct gaccccggat
     6601 gaatgccaac tgctgggcta tctggataaa ggcaaacgca aacgcaaaga aaaagcgggc
     6661 agcctgcaat gggcgtatat ggcgattgcg cgtctgggcg gctttatgga tagcaaacgt
     6721 accggcattg cgagctgggg tgcgctgtgg gaaggttggg aagcgctgca aagcaaactg
     6781 gatggctttc tggccgcgaa agacctgatg gcgcagggca ttaaaatcta aaagcttgat
     6841 gggggatcta atacgactca ctatagggag atttgataga ttaaaaagga aaggaggaaa
     6901 gaaataatgg ctcgtgggta atgactccaa cttattgata gtgttttatg ttcagataat
     6961 gcccgatgac tttgtcatgc agctccaccg attttgagaa cgacagcgac ttccgtccca
     7021 gccgtgccag gtgctgcctc agattcaggt tatgccgctc aattcgctgc gtatatcgct
     7081 tgctgattac gtgcagcttt cccttcaggc gggattcata cagcggccag ccatccgtca
     7141 tccatatcac cacgtcaaag ggtgacagca ggctcataag acgccccagc gtcgccatag
     7201 tgcgttcacc gaatacgtgc gcaacaaccg tcttccggag actgtcatac gcgtaaaaca
     7261 gccagcgctg gcgcgattta gccccgacat agccccactg ttcgtccatt tccgcgcaga
     7321 cgatgacgtc actgcccggc tgtatgcgcg aggttaccga ctgcggcctg agttttttaa
     7381 gtgacgtaaa atcgtgttga ggccaacgcc cataatgcgg gctgttgccc ggcatccaac
     7441 gccattcatg gccatatcaa tgattttctg gtgcgtaccg ggttgagaag cggtgtaagt
     7501 gaactgcagt tgccatgttt tacggcagtg agagcagaga tagcgctgat gtccggcggt
     7561 gcttttgccg ttacgcacca ccccgtcagt agctgaacag gagggacagc tgatagaaac
     7621 agaagccact ggagcacctc aaaaacacca tcatacacta aatcagtaag ttggcagcat
     7681 cacctggctc gtgtacagtt taaacaacgt gaatctactg acgcaatctt tgttcactgc
     7741 tcggctacca agccaagtca gaatgttggt gtccgtgaga ttcgccagtg gcacaaagag
     7801 cagggttggc tcgatgtggg ataccacttt atcatcaagc gagacggtac tgtggaggca
     7861 ggacgagatg agatggctgt aggctctcac gctaagggtt acaaccacaa ctctatcggc
     7921 gtctgccttg ttggtggtat cgacgataaa ggtaagttcg acgctaactt tacgccagcc
     7981 caaatgcaat cccttcgctc actgcttgtc acactgctgg ctaagtacga aggcgctgtg
     8041 cttcgcgccc atcatgaggt ggcgccgaag gcttgccctt cgttcgacct taagcgttgg
     8101 tgggagaaga acgaactggt cacttctgac cgtggataag atcccatggt acgcgtgcta
     8161 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     8221 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct aggggtatat
     8281 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggtg agcggaaatg
     8341 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     8401 agagggccgc ggcaaagccg tttttccata ggcttcgccc ccctgacaag catcacgaaa
     8461 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8521 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     8581 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagtcccgg gtaggcagtt
     8641 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     8701 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     8761 actg
//
