LOCUS       sequence-372970-        5148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobilisable shuttle and expression vector. Replicates in many 
            Gram-negative bacteria. Has multiple cloning site with blue/white 
            selection function. Cloned genes driven by derepressed lac 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5148)
  AUTHORS   Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM 2nd, 
            Peterson KM
  TITLE     Four new derivatives of the broad-host-range cloning vector 
            pBBR1MCS, carrying different antibiotic-resistance cassettes.
  JOURNAL   Gene. 1995 Dec 1;166(1):175-6.
  PUBMED    8529885
REFERENCE   2  (bases 1 to 5148)
  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: "Gene."; 
            date: "1995-12-1"; volume: "166(1)"; pages: "175-6"
COMMENT     Sequence Label: pBBR1MCS-2
FEATURES             Location/Qualifiers
     source          1..5148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1839..2633
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1893..1912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2503..2522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    3038..3059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3074..3079,3080..3097,3098..3104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3112..3128
                     /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     3117..3139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3136..3152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3136..3152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3148..3513
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     primer_bind     3171..3191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3173..3191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    3204..3311
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3220..3236
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3221..3237
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3271..3287)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3271..3287)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3319..3338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3320..3338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3348..3365)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3348..3364)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3357..3379)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4073..4735)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      join(4736..5148,1..359)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ggcgggcggg cttcgccctt cgactgcccc cactcgcata ggcttgggtc gttccaggcg
       61 cgtcaaggcc aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg
      121 tccaaccggc aagcgaagcg cgcaggccgc aggccggagg cttttcccca gagaaaatta
      181 aaaaaattga tggggcaagg ccgcaggccg cgcagttgga gccggtgggt atgtggtcga
      241 aggctgggta gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc
      301 atcagcttgt ccagcagggt tgtccacggg ccgagcgaag cgagccagcc ggtggccgct
      361 cgcggccatc gtccacatat ccacgggctg gcaagggagc gcagcgaccg cgcagggcga
      421 agcccggaga gcaagcccgt agggcgccgc agccgccgta ggcggtcacg actttgcgaa
      481 gcaaagtcta gtgagtatac tcaagcattg agtggcccgc cggaggcacc gccttgcgct
      541 gcccccgtcg agccggttgg acaccaaaag ggaggggcag gcatggcggc atacgcgatc
      601 atgcgatgca agaagctggc gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac
      661 cgcgagcgcg agacgcccaa cgctgacgcc agcaggacgc cagagaacga gcactgggcg
      721 gccagcagca ccgatgaagc gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc
      781 aaggacgctg tgttggcggt cgagtacgtc atgacggcca gcccggaatg gtggaagtcg
      841 gccagccaag aacagcaggc ggcgttcttc gagaaggcgc acaagtggct ggcggacaag
      901 tacggggcgg atcgcatcgt gacggccagc atccaccgtg acgaaaccag cccgcacatg
      961 accgcgttcg tggtgccgct gacgcaggac ggcaggctgt cggccaagga gttcatcggc
     1021 aacaaagcgc agatgacccg cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg
     1081 ctgcaacggg gcatcgaggg cagcaaggca cgtcacacgc gcattcaggc gttctacgag
     1141 gccctggagc ggccaccagt gggccacgtc accatcagcc cgcaagcggt cgagccacgc
     1201 gcctatgcac cgcagggatt ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg
     1261 gaagccgtgg ccgaccggct gacaaaagcg gttcggcagg ggtatgagcc tgccctacag
     1321 gccgccgcag gagcgcgtga gatgcgcaag aaggccgatc aagcccaaga gacggcccga
     1381 gaccttcggg agcgcctgaa gcccgttctg gacgccctgg ggccgttgaa tcgggatatg
     1441 caggccaagg ccgccgcgat catcaaggcc gtgggcgaaa agctgctgac ggaacagcgg
     1501 gaagtccagc gccagaaaca ggcccagcgc cagcaggaac gcgggcgcgc acatttcccc
     1561 gaaaagtgcc acctgggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     1621 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     1681 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     1741 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     1801 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     1861 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1921 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1981 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2041 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2101 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2161 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2221 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2281 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2341 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2401 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2461 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2521 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2581 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     2641 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     2701 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     2761 gatcctccag cgcggggatc tcatgctgga gttcttcgcc cacccccatg ggcaaatatt
     2821 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     2881 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtttttatgc atgcgcccaa
     2941 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3001 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     3061 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     3121 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     3181 ctcactaaag ggaacaaaag ctgggtaccg ggccccccct cgaggtcgac ggtatcgata
     3241 agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg gccgccaccg
     3301 cggtggagct ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     3361 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3421 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3481 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     3541 taaatttttg ttaaatcagc tcatttttta accaataggc cgtactgcga tgagtggcag
     3601 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc
     3661 tgaataagtg ataataagcg gatgaatggc agaaattcga aagcaaattc gacccggtcg
     3721 tcggttcagg gcagggtcgt taaatagccg cttatgtcta ttgctggttt accggtttat
     3781 tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag tttgctcagg
     3841 ctctccccgt ggaggtaata attgacgata tgatcattta ttctgcctcc cagagcctga
     3901 taaaaacggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
     3961 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgaggcggct
     4021 acagccgata gtctggaaca gcgcacttac gggttgctgc gcaacccaag tgctaccggc
     4081 gcggcagcgt gacccgtgtc ggcggctcca acggctcgcc atcgtccaga aaacacggct
     4141 catcgggcat cggcaggcgc tgctgcccgc gccgttccca ttcctccgtt tcggtcaagg
     4201 ctggcaggtc tggttccatg cccggaatgc cgggctggct gggcggctcc tcgccggggc
     4261 cggtcggtag ttgctgctcg cccggataca gggtcgggat gcggcgcagg tcgccatgcc
     4321 ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac ggcggcactg aacaccgaca
     4381 ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc attgaccacg taggccgaca
     4441 cggtgccggg gccgttgagc ttcacgacgg agatccagcg ctcggccacc aagtccttga
     4501 ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt ggaaagtgtc ttctggctga
     4561 ccaccacggc gttctggtgg cccatctgcg ccacgaggtg atgcagcagc attgccgccg
     4621 tgggtttcct cgcaataagc ccggcccacg cctcatgcgc tttgcgttcc gtttgcaccc
     4681 agtgaccggg cttgttcttg gcttgaatgc cgatttctct ggactgcgtg gccatgctta
     4741 tctccatgcg gtaggggtgc cgcacggttg cggcaccatg cgcaatcagc tgcaactttt
     4801 cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca gtcaattaca gattttcttt
     4861 aacctacgca atgagctatt gcggggggtg ccgcaatgag ctgttgcgta cccccctttt
     4921 ttaagttgtt gatttttaag tctttcgcat ttcgccctat atctagttct ttggtgccca
     4981 aagaagggca cccctgcggg gttcccccac gccttcggcg cggctccccc tccggcaaaa
     5041 agtggcccct ccggggcttg ttgatcgact gcgcggcctt cggccttgcc caaggtggcg
     5101 ctgccccctt ggaacccccg cactcgccgc cgtgaggctc ggggggca
//
