LOCUS       sequence-409802-        8124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniICE-tet(M)cat(PC194)_PT7-GFP; used to integrate into JAB1000 B. 
            subtilis XPORT strain with IPTG-inducible GFP under T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8124)
  AUTHORS   Brophy JAN, Triassi AJ, Adams BL, Renberg RL, Stratis-Cullum DN, 
            Grossman AD, Voigt CA
  TITLE     Engineered integrative and conjugative elements for efficient and 
            inducible DNA transfer to undomesticated bacteria.
  JOURNAL   Nat Microbiol. 2018 Sep;3(9):1043-1053. doi: 
            10.1038/s41564-018-0216-5. Epub 2018 Aug 20.
  PUBMED    30127494
REFERENCE   2  (bases 1 to 8124)
  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: 
            "10.1038/s41564-018-0216-5"; journalName: "Nat Microbiol"; date: 
            "2018-09"; volume: "3"; issue: "9"; pages: "1043-1053"
COMMENT     Sequence Label: pJAB988
FEATURES             Location/Qualifiers
     source          1..8124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(86..877,878..946))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     709..728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(947..1051)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2301..2320
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2301..2319
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2356..3072
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(2383..2411)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3006..3026
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      7000..7086
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7105..7132
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      7212..7246
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(7451..8039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7531..7550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
       61 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      121 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      181 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      241 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      301 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      361 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
      421 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      481 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      541 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
      601 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
      661 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
      721 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
      781 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
      841 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
      901 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
      961 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1021 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgagg
     1081 tgttttgata gaaatgtccg gtcaaggttg ccgtcaattt gaatccttct tggaatgccg
     1141 gaagaaaaca tggtacgact tctttcaaga ttgcatgcaa caaggcggtt cgttcactcg
     1201 ctttgattta gctattgatg ataaaaagac atacttttct ataccagaac tgttaaagaa
     1261 agcccaaaag ggcgaatgta tttcacgatt tagaaaatct gatttcaatg gctcctttga
     1321 tctatctgat ggcattacag gcgggacgac aatatatttc ggttctaaaa agtcagaagc
     1381 gtacctttgc ttttatgaga aaaactatga acaagcagaa aagtataaca ttccgttaga
     1441 agaactaggc gattggaatc gttatgaatt gcggcttaaa aatgaacgtg cacaagtggc
     1501 gatagatgct ttattgaaaa caaaggattt aaccttaatc gctatgcaga tcattaataa
     1561 ctatgtaagg ttcgttgatg ctgatgaaaa catcactaga gagcattgga aaacgagcct
     1621 tttttggtct gattttatag gcgatgttgg gagacttcct ttgtacgtca aaccacaaaa
     1681 agacttctat cagaaatcaa gaaactggct tcgtaactct tgtgctccca ctatgaaaat
     1741 ggtgcttgaa gctgatgaac atttagggaa aaccgatctg tcagatatga ttgcagaagc
     1801 tgaactggct gacaaacata aaaaaatgtt agatgtgtat atggctgatg ttgctgacat
     1861 ggtcgtttaa aaccaagctg ctaaacctta ataaacacag gtgatgcgat taaacgtatg
     1921 tgtcaacggt accggaacga agtgaggata aggagttgac atattcagcg ctacccaaac
     1981 cctcttggca ttgaacaaac aaccgtttgt tcctgcctac cggcgaggga gtcccccgat
     2041 gattcggggg aataggggga tagattaaca gcaatccaac gagctaccgt tggtgggttt
     2101 gggtgaaacc caagaactaa cttggaggga taagactagt gcctcctcat cctcttcatc
     2161 ctcttcgtct tggtagcttt ttaaatatgg cgcttcatag agtaattctg taaaggtcca
     2221 attctcgttt tcatacctcg gtataatctt acctatcacc tcaaatggtt ctctagagtc
     2281 gacgaattca acagctagag taatacgact cactataggg agatgtttga gtcggctgaa
     2341 aggtggtgaa ctactatgaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     2401 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     2461 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     2521 tggccaacac ttgtcactac tttcgcgtat ggtcttcaat gctttgcgag atacccagat
     2581 catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
     2641 actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt
     2701 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
     2761 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
     2821 caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt
     2881 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     2941 gacaaccatt acctgtccac acaatctaag ctttcgaaag atcccaacga aaagagagac
     3001 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta
     3061 tacaaatact agcctgcagg aagaactaac ttggagggat aaggatttcc tccgacctaa
     3121 tttaatccat tgtttgtaaa catttcatat ctaattatat aacatagact cctattctaa
     3181 ctacttattt taaagcagtc tggattgttt gggtaattca taaaaaaata aaagaaagaa
     3241 ggaggaatag caaatattgg tacattatta cagctatttt gtaatcacgt actctctttg
     3301 ataaaaaatt ggagattcct ttacaaatat gctcttacgt gctattattt aagtatctat
     3361 ttaaaaggag ttaataaata tgcggcaagg tattattaaa taaactgtca atttgatagc
     3421 gggaacaaat aattggatgt ccttttttag gagggcttag ttttttgtac ccagtttaag
     3481 aataccttta tcatgtgatt ctaaagtatc cggagaatat ctgtatgctt tgtatgccta
     3541 tggttatgca taaaaatccc agtgataaga gtatttatca ctgggatttt tatgcccttt
     3601 tgggcttttg aatggaggaa aatcacatga aaattattaa tattggagtt ttagctcatg
     3661 ttgatgcagg aaaaactacc ttaacagaaa gcttattata taacagtgga gcgattacag
     3721 aattaggaag cgtggacaaa ggtacaacga ggacggataa tacgctttta gaacgtcaga
     3781 gaggaattac aattcagaca ggaataacct cttttcagtg ggaaaatacg aaggtgaaca
     3841 tcatagacac gccaggacat atggatttct tagcagaagt atatcgttca ttatcagttt
     3901 tagatggggc aattctactg atttctgcaa aagatggcgt acaagcacaa actcgtatat
     3961 tatttcatgc acttaggaaa atggggattc ccacaatctt ttttatcaat aagattgacc
     4021 aaaatggaat tgatttatca acggtttatc aggatattaa agagaaactt tctgccgaaa
     4081 ttgtaatcaa acagaaggta gaactgtatc ctaatgtgtg tgtgacgaac tttaccgaat
     4141 ctgaacaatg ggatacggta atagagggaa acgatgacct tttagagaaa tatatgtccg
     4201 gtaaatcatt agaagcattg gaactcgaac aagaggaaag cataagattt cagaattgtt
     4261 ctctgttccc tctttatcat ggaagtgcaa aaagtaatat agggattgat aaccttatag
     4321 aagttattac taataaattt tattcatcaa cacatcgagg tccgtctgaa ctttgcggaa
     4381 atgttttcaa aattgaatat acaaaaaaaa gacaacgtct tgcatatata cgcctttata
     4441 gtggagtact acatttacga gattcggtta gagtatcaga aaaagaaaaa ataaaagtta
     4501 cagaaatgta tacttcaata aatggtgaat tatgtaagat tgatagagct tattctggag
     4561 aaattgttat tttgcaaaat gagtttttga agttaaatag tgttcttgga gatacaaaac
     4621 tattgccaca gagaaaaaag attgaaaatc cgcaccctct actacaaaca actgttgaac
     4681 cgagtaaacc tgaacagaga gaaatgttgc ttgatgccct tttggaaatc tcagatagtg
     4741 atccgcttct acgatattac gtggattcta cgacacatga aattatactt tctttcttag
     4801 ggaaagtaca aatggaagtg attagtgcac tgttgcaaga aaagtatcat gtggagatag
     4861 aactaaaaga gcctacagtc atttatatgg agagaccgtt aaaaaatgca gaatatacca
     4921 ttcacatcga agtgccgcca aatcctttct gggcttccat tggtttatct gtatcaccgc
     4981 ttccgttggg aagtggaatg cagtatgaga gctcggtttc tcttggatac ttaaatcaat
     5041 catttcaaaa tgcagttatg gaagggatac gctatggttg tgaacaagga ttgtatggtt
     5101 ggaatgtgac ggactgtaaa atctgtttta agtatggctt atactatagc cctgttagta
     5161 ccccagcaga ttttcggatg cttgctccta ttgtattgga acaagtctta aaaaaagctg
     5221 gaacagaatt gttagagcca tatcttagtt ttaaaattta tgcgccacag gaatatcttt
     5281 cacgagcata caacgatgct cctaaatatt gtgcgaacat cgtagacact caattgaaaa
     5341 ataatgaggt cattcttagt ggagaaatcc ctgctcggtg tattcaagaa tatcgtagtg
     5401 atttaacttt ctttacaaat ggacgtagtg tttgtttaac agagttaaaa gggtaccatg
     5461 ttactaccgg tgaacctgtt tgccagcccc gtcgtccaaa tagtcggata gataaagtac
     5521 gatatatgtt caataaaata acttagtgta ttttatgttg ttatataaat atggtttctt
     5581 gttaaataag atgaaatatt ttttaataag atttgaatta aagtgtccgg caatagttac
     5641 ccttattatc aagataagaa agaaaaggat ttttcgctac gctcaaatcc tttaaaaaaa
     5701 cacaaaagac cacatttttt aatgtggtct tttattcttc aactaaagca cccattagtt
     5761 caacaaacga aaattggata aagtgggata tttttaaaat atatatttat gttacagtaa
     5821 tattgacttt taaaaaagga ttgattctaa tgaagaaagc agacaagtaa gcctcctaaa
     5881 ttcactttag ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttgatttaga
     5941 caattggaag agaaaagaga tatttaatca ttatttgaac caacaaacga cttttagtat
     6001 aaccacagaa attgatatta gtgttttata ccgaaacata aaacaagaag gatataaatt
     6061 ttaccctgca tttattttct tagtgacaag ggtgataaac tcaaatacag cttttagaac
     6121 tggttacaat agcgacggag agttaggtta ttgggataag ttagagccac tttatacaat
     6181 ttttgatggt gtatctaaaa cattctctgg tatttggact cctgtaaaga atgacttcaa
     6241 agagttttat gatttatacc tttctgatgt agagaaatat aatggttcgg ggaaattgtt
     6301 tcccaaaaca cctatacctg aaaatgcttt ttctctttct attattccat ggacttcatt
     6361 tactgggttt aacttaaata tcaataataa tagtaattac cttctaccca ttattacagc
     6421 aggaaaattc attaataaag gtaattcaat atatttaccg ctatctttac aggtacatca
     6481 ttctgtttgt gatggttatc atgcaggatt gtttatgaac tctattcagg aattgtcaga
     6541 taggcctaat gactggcttt tataatatga gataatgccg actgtacttt ttacagtcgg
     6601 tgttctaatg tcactaacct gccttagata attggaagaa gtacatccgc aactgtccat
     6661 actctgatgt ctagaggatc cccgggtacc gagctcgaat tcgctatttt atatccatag
     6721 ttgttaatta aataaactta atttagttta tttatagatt tcattggctt ctaaattttt
     6781 tatctagata ataattattt tagttaattt tattctagat tatatatgat atgatctttc
     6841 atttccataa aactaaagta agtgtaaacc tattcattgt tttaaaaata tctcttgcca
     6901 gtcacgttac gttattagtt atagttatta taacatgtat tcacgaacga aaatcgccat
     6961 tcgccagctg caggaattcg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     7021 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     7081 acaaatccgc cgctctagct aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     7141 tacaaactct tgttaactct agagctgcct gccgcgtttc ggtgatgaag atcttcccga
     7201 tgattaatta attcagaacg ctcggttgcc gccgggcgtt ttttatgcag caatggcaag
     7261 aacgttgctc gagggtaaat gtgagcactc acaattcatt ttgcaaaagt atttagaaag
     7321 attactatat cctcaaacag gcggtaaccg gcctcttcat cgggaatgcg cgcgaccttc
     7381 aacatcgccg gcatgtcccc ctggcggacg ggaagtatcc agctcgaggt cgggccgcgt
     7441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7861 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8101 ggattttggt catgagatta tcaa
//
