LOCUS       sequence-503497-        7260 bp    DNA     circular SYN 19-MAR-2026
DEFINITION  aTC-Inducible P21 with 1x TetO site driving mScarlet | pTHT15 Ori | 
            Erythromycin Resistance Cassette | Conjugative Vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7260)
  AUTHORS   Ortiz L, Esmurria A, Gilbert C, Crits-Christoph A, Barnum TP, 
            Mancuso CP, Kang SC, Leung J, Fenn K, Abrams MB, Brumwell SL, Lee 
            HH, Ostrov N
  TITLE     A genetic platform for a biocementation bacterium
  JOURNAL   bioRxiv 2025.12.05.692644
REFERENCE   2  (bases 1 to 7260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.12.05.692644"
COMMENT     Sequence Label: pGLU_511_71
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            909..1017
                     /note="incP origin of transfer"
     primer_bind     complement(1171..1187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1171..1187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1184..1206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      1377..1765
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(1914..2008)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2029..2688)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2603..2622
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2689..2791)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2878..2900
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2892..2909
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2893..2909
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3251..4255)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     primer_bind     complement(4776..4793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(6391..7017)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     6922..6941
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    7187..7205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             7227..7244
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tggtgagtaa gggtgaagcc gtgattaaag agttcatgcg cttcaaagtg catatggagg
       61 gctcaatgaa cggccatgaa tttgaaattg aaggtgaagg agaaggccgt ccgtatgagg
      121 gcacccagac cgccaaactg aaagtcacaa agggtggccc gctgccgttt tcgtgggata
      181 ttttatcccc tcaattcatg tacggcagcc gggcgtttat taaacacccg gcggatatcc
      241 cggattacta taagcaatct tttccagaag gctttaaatg ggagcgcgtg atgaattttg
      301 aagacggagg tgctgttacc gtgacccagg acacgagctt agaagacggt accttgatct
      361 ataaggtaaa gctgcgtggt accaatttcc cgccggatgg gccagttatg cagaaaaaga
      421 ctatgggctg ggaagcatcg acggagcgtc tttaccccga agacggcgtc ctgaaaggcg
      481 atattaaaat ggccctgcga ctgaaagacg gcggtcgcta ccttgcagat ttcaaaacaa
      541 cgtataaagc gaaaaaacct gtgcagatgc ccggggcgta caacgttgat cgcaaactag
      601 atatcacttc acacaacgaa gattacacgg tagtcgaaca gtatgaacgt agcgaggggc
      661 gccatagcac cggaggcatg gatgaactct ataaataagc ttcgctcgtg ctcgccgcga
      721 ccttcctctt catgcacctc ttcggcatcg ggctgcacaa cgctacagag gagcttttcc
      781 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
      841 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
      901 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
      961 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtac
     1021 gctaggtata cttgagacca ccccaggata catagattac cacaactccg agcccttcca
     1081 cctagcgtgc taaccacgca cgctattgtt gtataggcta ggtggaggct cagtgatgat
     1141 aagtctgcga tggtggatgc atgtgtcatg gtcatagctg tttcctgtgt gaaattgtta
     1201 tccgctcaga gggcacaatc ctattccgcg ctatccgaca atctccaaga cattaggtgg
     1261 agttcagttc ggcgtatggc atatgtcgct ggaaagaaca tgtgagcaaa aggccagcaa
     1321 aaggccagga accgtaaaaa ggccgcgttg ctggcgttac agctgggcgc gccccatgtc
     1381 agccgttaag tgttcctgtg tcactcaaaa ttgctttgag aggctctaag ggcttctcag
     1441 tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaggctt taaaagcctt
     1501 atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt
     1561 atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     1621 ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt
     1681 agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac
     1741 aggttgaact gctgatcttc agatcggcgc gccggccggc cgaagatcct ttgatctttt
     1801 ctacggggtc tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag cgtaaatggg
     1861 tagggggctt caaatcgtcc ccccatacga tataagttgt tactagtgct tggattctca
     1921 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     1981 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     2041 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     2101 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     2161 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     2221 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     2281 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     2341 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     2401 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     2461 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     2521 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     2581 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     2641 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     2701 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     2761 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tgatacctga aacaaaaccc
     2821 atcgtacggc caaggaagtc tccaataact gtgatccacc acaagcgcca gggttttccc
     2881 agtcacgacg ttgtaaaacg acggccagtc atgcataatc cgcacgcatc tggaataagg
     2941 aagtgccatt ccgcctgacc ttagcgtgcg aacagcacgc tattgttgta tcattactcg
     3001 catccattct caggctgtct cgtctcgtct ccgaggtctc aggagaatga gtcagggtca
     3061 cgcgtaggac gcgtcgagta gggtttcttg tcgactccat gtcagttccg cagccacacg
     3121 ccgtgtgaag ctggggaggt tcatttgata tgcctcctaa atttttatct aaagtgaatt
     3181 taggaggctt acttgtctgc tttcttcatt agaatcaatc cttttttaaa agtcaatccc
     3241 gtttgttgaa ctactcttta ataaaataat ttttccgttc ccaattccac attgcaataa
     3301 tagaaaatcc atcttcatcg gctttttcgt catcatctgt atgaatcaaa tcgccttctt
     3361 ctgtgtcatc aaggtttaat tttttatgta tttcttttaa caaaccacca taggagatta
     3421 accttttacg gtgtaaacct tcctccaaat cagacaaacg tttcaaattc ttttcttcat
     3481 catcggtcat aaaatccgta tcctttacag gatattttgc agtttcgtca attgccgatt
     3541 gtatatccga tttatattta tttttcggtc gaatcatttg aacttttaca tttggatcat
     3601 agtctaattt cattgccttt ttccaaaatt gaatccattg tttttgattc acgtagtttt
     3661 ctgtattctt aaaataagtt ggttccacac ataccaatac atgcatgtgc tgattataag
     3721 aattatcttt attatttatt gtcacttccg ttgcacgcat aaaaccaaca agatttttat
     3781 taattttttt atattgcatc attcggcgaa atccttgagc catatctgac aaactcttat
     3841 ttaattcttc gccatcataa acatttttaa ctgttaatgt gagaaacaac caacgaactg
     3901 ttggcttttg tttaataact tcagcaacaa ccttttgtga ctgaatgcca tgtttcattg
     3961 ctctcctcca gttgcacatt ggacaaagcc tggatttaca aaaccacact cgatacaact
     4021 ttctttcgcc tgtttcacga ttttgtttat actctaatat ttcagcacaa tcttttactc
     4081 tttcagcctt tttaaattca agaatatgca gaagttcaaa gtaatcaaca ttagcgattt
     4141 tcttttctct ccatggcctc acttttccac tttttgtctt gtccactaaa acccttgatt
     4201 tttcatctga ataaatgcta ctattaggac acataatatt aaaagaaacc cccatctatt
     4261 tagttatttg tttggtcact tataacttta acagatgggg tttttctgtg caaccaattt
     4321 taagggtttt caatacttta aaacacatac ataccaacac ttcaacgcac ctttcagcaa
     4381 ctaaaataaa aatgacgtta tttctatatg tatcaagaat agaaagaact cgtttttcgc
     4441 tacgctcaaa acgcaaaaaa agcactcatt cgagtgcttt ttcttatcgc tccaaatcat
     4501 gcgatttttt cctctttgct tttctttgct cacgaagttc tcgatcacgc tgcaaaacat
     4561 cttgaagcga aaaagtattc ttcttttctt ccgatcgctc atgctgacgc acgaaaagcc
     4621 ctctaggcgc ataggaacaa ctcctaaatg catgtgaggg gttttctcgt ccatgtgaac
     4681 agtcgcatac gcaatatttt gtttcccata ctgcattaat gaatcggcca acgcgcgggg
     4741 agaggcggtt tgcgtattgg gcgctcttgc gcttcctcgc tcactgactc gctgcgctcg
     4801 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4861 gaatcagggg cgctagcaag gaggatcctt taactctggc aaccctcaaa attgaatgag
     4921 acatgctaca cctccggata ataaatatat ataaacgtat atagatttca taaagtctaa
     4981 cacactagac ttatttactt cgtaattaag tcgttaaacc gtgtgctcta cgaccaaaac
     5041 tataaaacct ttaagaactt tcttttttta caagaaaaaa gaaattagat aaatctctca
     5101 tatcttttat tcaataatcg catccgattg cagtataaat ttaacgatca ctcatcatgt
     5161 tcatatttat cagaggtcgt gctataatta tactaatttt ataaggagga aaaaatatgg
     5221 gcatttttag tatttttgta atcagcacag ttcattatca accaaacaaa aaataagtgg
     5281 ttataatgaa tcgttaataa gcaaaattca tataaccaaa ttaaagaggg ttataaatga
     5341 acgagaaaaa tataaaacac agtcaaaact ttattacttc aaaacataat atagataaaa
     5401 taatgacaaa tataagatta aatgaacatg ataatatctt tgaaatcggc tcaggaaaag
     5461 gccattttac ccttgaatta gtaaagaggt gtaatttcgt aactgccatt gaaatagacc
     5521 ataaattatg caaaactaca gaaaataaac ttgttgatca cgataatttc caagttttaa
     5581 acaaggatat attgcagttt aaatttccta aaaaccaatc ctataaaata tatggtaata
     5641 taccttataa cataagtacg gatataatac gcaaaattgt ttttgatagt atagctaatg
     5701 agatttattt aatcgtggaa tacgggtttg ctaaaagatt attaaataca aaacgctcat
     5761 tggcattact tttaatggca gaagttgata tttctatatt aagtatggtt ccaagagaat
     5821 attttcatcc taaacctaaa gtgaatagct cacttatcag attaagtaga aaaaaatcaa
     5881 gaatatcaca caaagataaa caaaagtata attatttcgt tatgaaatgg gttaacaaag
     5941 aatacaagaa aatatttaca aaaaatcaat ttaacaattc cttaaaacat gcaggaattg
     6001 acgatttaaa caatattagc tttgaacaat tcttatctct tttcaatagc tataaattat
     6061 ttaataagta agcttgttaa gggatgcata aactgcatcc cttaacttgt ttttcgtgtg
     6121 cctatttttt gtgaatcgat tatgtctttt gcgcagtcgg ccgctatacc ctcgtctcaa
     6181 ccaaagcaat caacccatca accacctggt cgttgggatt gtcctgaccg aagcgggaca
     6241 ggtccacgag caggcaatgc ttgttgggca tggtgaggtg gatgtctttc acttccggca
     6301 cgcagccgag gactttttgc gccatatcgt aaagtgtgtg ctgaacggat tcgctttcgt
     6361 gttcagcgaa agaggcgaga aggagattcc ttaagaccca ctttcacatt taagttgttt
     6421 ttctaatccg catatgatca attcaaggcc gaataagaag gctggctctg caccttggtg
     6481 atcaaataat tcgatagctt gtcgtaataa tggcggcata ctatcagtag taggtgtttc
     6541 cctttcttct ttagcgactt gatgctcttg atcttccaat acgcaaccta aagtaaaatg
     6601 ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa
     6661 ggctaattga ttttcgagag tttcatactg tttttctgta ggccgtgtac ctaaatgtac
     6721 ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt tattacgtaa
     6781 aaaatcttgc cagctttccc cttctaaagg gcaaaagtga gtatggtgcc tatctaacat
     6841 ctcaatggct aaggcgtcga gcaaagcccg cttatttttt acatgccaat acaatgtagg
     6901 ctgctctaca cctagcttct gggcgagttt acgggttgtt aaaccttcga ttccgacctc
     6961 attaagcagc tctaatgcgc tgttaatcac tttactttta tctaatctag acatcatact
     7021 taaaacatcc ttccagatta tcttctgtta gtatacacaa aaaaagatga tgatgcccta
     7081 aaagagcacc atcaccgtat gagtttttca aaatctttcg acgcattccg aactattttt
     7141 ctatttgccg aagtattgac tttatttcaa tagatatgta gtatagtccc tatcagtgat
     7201 agagaaggag gtttttaatg ggttctcacc accaccacca tcacactacg agtacaggga
//
