LOCUS       sequence-463766-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Broad host range plasmid 
            with stability. L1-pBBR1-Tc-par-ELT4; GG assembly from pOGG011, 
            pOGG012, pOGG014, pOGG042, and pOGG004, TcR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  AUTHORS   Geddes BA, Williamson R, Schumacher J, Ardi A, Levin G, Cervenka E, 
            Huang R, diCenzo G
  TITLE     BEVA2.0: modular assembly of golden gate-compatible vectors with 
            expanded utility for genetic engineering.
  JOURNAL   Can J Microbiol. 2025 Jan 1;71:1-13. doi: 10.1139/cjm-2024-0246.
  PUBMED    39919271
REFERENCE   2  (bases 1 to 6972)
  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: "Can J 
            Microbiol."; date: "2025-01-1"; volume: "71:1-13. doi"; pages: " 
            10.1139/cjm-2024-0246"
COMMENT     Sequence Label: pNDGG004
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            140..248
                     /note="incP origin of transfer"
     CDS             complement(259..921)
                     /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      922..1693
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(4202..4288)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    4376..4397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4412..4417,4418..4435,4436..4442)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4450..4466
                     /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     4455..4477
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4474..4490
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4474..4490
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4486..4809
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    4503..4559
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4560..4577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4560..4576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4785..4804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      4914..5008
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5017..5667)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             5773..6972
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 actacttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
       61 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
      121 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
      181 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
      241 gctggccgta ggccggccct accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg
      301 ctcgccatcg tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg
      361 ttcccattcc tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg
      421 ctggctgggc ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt
      481 cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac
      541 caccacggcg gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac
      601 gcggtcattg accacgtagg ccgacacggt gccggggccg ttgagcttca cgacggagat
      661 ccagcgctcg gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat
      721 gagcttggaa agagtctttt ggctgaccac cacggcgttc tggtggccca tctgcgcgac
      781 gaggtgatgc agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc
      841 atgcgctttg cgttccgttt ggacccagtg accgggcttg ttcttggctt gaatgccgat
      901 ttctctggac tgcgtggcca tgcttatctc catgcggtag gggtgccgca cggttgcggc
      961 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa
     1021 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc
     1081 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg
     1141 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct
     1201 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc
     1261 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg
     1321 aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg cccccactcg cataggcttg
     1381 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg
     1441 ccttcctctt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc
     1501 cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt
     1561 gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca
     1621 ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccgagc gaagcgagcc
     1681 agccggtggc cgctcgcggc catcgtccac atatccacgg gctggcaagg gagcgcagcg
     1741 accgcgcagg gcgaagcccg gagagcaagc ccgtaggggg ttactgcatg agcttgtgga
     1801 agtgtgctga gcataaatga ttctcctagc tgttctttgg gtaagtacgc catcaggacg
     1861 ttgtgagtgg cgcgattttt agcggctgaa atcagccctt gagcctgtcg gcaagtcgcg
     1921 tcatgaggtc catgcgctca tgcaggatcg ccacgaccaa cgcgggttcg cccgcacgcg
     1981 gcaggcaaaa aacgtagtgg tgttcgcagc gggccatccg cagcgcggga aagagttcgc
     2041 tcatgtcctt aaacgggcct tcgccggcgg caagcctggc tatgccctgt tccagcttag
     2101 cgatatagcg gcgcacctgc gccgcgcccc actcccggcg cgtgtagcgg atgatgccgc
     2161 gtagatcggc ttcggcctca gccgtgagga tgtaggccgt caagcgcgat ccccgctgag
     2221 ttcttcatca agaatttcgc cgacgctctt ggtggacacc ttgccggcaa gcccatcgtt
     2281 gatgcggttc cccagcatgg ttttcagttc ctgccatgcc tgatcggcat cagcgtcacc
     2341 ggggaacaga cgttcgaggg cgtattgctt aatggtcttg ccctgcaagg cggccagggc
     2401 tttcaggctc tggtgctgct ggtccgtcat gtcgattgtc aggcggctca ttggataacc
     2461 tccataaaat acacgtaacc acattagcac atatgtgggc gtgaggctac agcgcgaggc
     2521 gcattaaggt cgggaaaatg cgctaggcgc atttaaattg cgtattgctg taatgcgcca
     2581 tgccggctag actaggccca aatgggtata cccaatttga ccaaggggga cgcgatgagg
     2641 gcggccaagc actaccgaca acttctatcc atcgacttca acatcgaggc gctggccttc
     2701 gtgcctggac ccgacggcac acgcggccgg cgcatccacg tcctggggcg cgaggtccgc
     2761 gaccggcccg gcctggtcga gtacctttcg ccggcgttcg gctcgcgggt ggcgctggac
     2821 ggctactgca aggccaattt cgatgcagtg ctgcacctgg cgtaccccga tcatcagcaa
     2881 tggggccacg catgaagcgc cgaagctacg ccatgctgcg cgccgctgcc gcgctggccg
     2941 tcctggtcgt tgcctcgccg gcatgggccg agctgcgcgg cgaggtcgtg cgcatcatcg
     3001 acggcgacac catcgacgtg ctggtagaca agcagccggt gcgcgtgcgc ctggtggaca
     3061 ttgacgcgcc ggaaaagcgg caagccttcg gcgaacgtgc gcgccaggcg ctggccggca
     3121 tggtgttccg ccggcacgtc ctggtcgacg agaaggacac cgaccgttac ggccgcacgc
     3181 tgggcaccgt ctgggtcaac atggagctgg ccagccggcc gccgcagccg cgcaacgtca
     3241 acgccgcgat ggttcaccag ggcatggcgt gggcctatcg cttccacggc cgcgcggccg
     3301 accctgaaat gctgcggctc gaacaggagg cgcgaggcaa gcgcgtcggc ctctggtccg
     3361 atccgcacgc cgtcgagccg tggaaatggc gacgcgagag caacaaccgg agggacgaag
     3421 gttgaaggtc gcccgcatct acctgcgcgc cagtacggac gagcagaatc ttgaacgcca
     3481 ggagagcctt gtagcggcca cgcgggccgc cgggtactac gtcgccggca tctaccgcga
     3541 gaaggcgtcc ggcgcacgcg ccgaccggcc cgagctgctg cgcatgatcg cggacctgca
     3601 acctggtgaa gtcgtcgttg cggagaagat cgaccgcatc agccgcttgc cgttggccga
     3661 ggccgagcgc ctggttgcgt cgatccgggc caaaggggcc aagctggccg tgcctggcgt
     3721 ggtggacctg tcggagctgg ccgccgaggc gaacggagtg gcgaaaatcg ttctggaatc
     3781 cgtccaggac atgcttttga agctcgcctt gcagatggcc cgcgacgact acgaggatcg
     3841 gcgcgagcgt caacgtcagg gtgtccagtt ggcgaaggcc gccggccgct acaccggccg
     3901 caaacgtgac gccggcatgc acgaccgcat catcacgctt cgctccggcg gatcgagcat
     3961 tgccaagacg gccaagctgg tcggatgcag cccgagccag gtcaaacgag tgtgggcggc
     4021 ctggaacgcg cagcagcaaa aataaagccg ggcagtgccc ggcttttctc accttttcgc
     4081 gtcccgcagg gccgctgcga gcgccctacc tagatcctcg ctttccccct cggtgtagtc
     4141 cggccagggc acgaagggcg cggatgcgaa cctgttgagc aggtcagatc tagggcggcg
     4201 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     4261 ctttcgactg agcctttcgt tttatttgat gcctgccgaa ttcggatccg gagtgagacc
     4321 gcagctggca cgacaggttt gccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4381 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4441 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4501 ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca
     4561 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4621 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4681 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     4741 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4801 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4861 tgtctgctcc cggcatccgc ttacagacaa gctgtgacgg tctcacgctc ttggactcct
     4921 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     4981 ccgccgggcg ttttttattg gtgagaatcc aggcaatcaa tcgtcaccct ttctcggtcc
     5041 ttcaacgttc ctgacaacga gcctcctttt cgccaatcca tcgacaatca ccgcgagtcc
     5101 ctgctcgaac gctgcgtccg gaccggcttc gtcgaaggcg tctatcgcgg cccgcaacag
     5161 cggcgagagc ggagcctgtt caacggtgcc gccgcgctcg ccggcatcgc tgtcgccggc
     5221 ctgctcctca agcacggccc caacagtgaa gtagctgatt gtcatcagcg cattgacggc
     5281 gtccccggcc gaaaaacccg cctcgcagag gaagcgaagc tgcgcgtcgg ccgtttccat
     5341 ctgcggtgcg cccggtcgcg tgccggcatg gatgcgcgcg ccatcgcggt aggcgagcag
     5401 cgcctgcctg aagctgcggg cattcccgat cagaaatgag cgccagtcgt cgtcggctct
     5461 cggcaccgaa tgcgtatgat tctccgccag catggcttcg gccagtgcgt cgagcagcgc
     5521 ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga acccccaacc gttccgccag
     5581 tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac aggtccaggg cggcacggat
     5641 cactgtattc ggctgcaact ttgtcatgct tgacacttta tcactgataa acataatatg
     5701 tccaccaact tatcagtgat aaagaatccg cgcgttcaat cggaccagcg gaggctggtc
     5761 cggaggccag acgtgaaacc caacataccc ctgatcgtaa ttctgagcac tgtcgcgctc
     5821 gacgctgtcg gcatcggcct gattatgccg gtgctgccgg gcctcctgcg cgatctggtt
     5881 cactcgaacg acgtcaccgc ccactatggc attctgctgg cgctgtatgc gttggtgcaa
     5941 tttgcctgcg cacctgtgct gggcgcgctg tcggatcgtt tcgggcggcg gccaatcttg
     6001 ctcgtgtcgc tggccggcgc cactgtcgac tacgccatca tggcgacagc gcctttcctt
     6061 tgggttctct atatcgggcg gatcgtggcc ggcatcaccg gggcgactgg ggcggtagcc
     6121 ggcgcttata ttgccgatat cactgatggc gatgagcgcg cgcggcactt cggcttcatg
     6181 agcgcctgtt tcgggttcgg gatggtcgcg ggacctgtgc tcggtgggct gatgggcggt
     6241 ttctcccccc acgctccgtt cttcgccgcg gcagccttga acggcctcaa tttcctgacg
     6301 ggctgtttcc ttttgccgga gtcgcacaaa ggcgaacgcc ggccgttacg ccgggaggct
     6361 ctcaacccgc tcgcttcgtt ccggtgggcc cggggcatga ccgtcgtcgc cgccctgatg
     6421 gcggtgttct tcatcatgca acttgtcgga caggtgccgg ccgcgctttg ggtcattttc
     6481 ggcgaggatc gctttcactg ggacgcgacc acgatcggca tttcgcttgc cgcatttggc
     6541 attctgcatt cactcgccca ggcaatgatc accggccctg tagccgcccg gctcggcgaa
     6601 aggcgggcac tcatgctcgg aatgattgcc gacggcacag gctacatcct gcttgccttc
     6661 gcgacacggg gatggatggc gttcccgatc atggtcctgc ttgcttcggg tggcatcgga
     6721 atgccggcgc tgcaagcaat gttgtccagg caggtggatg aggaacgtca ggggcagctg
     6781 caaggctcac tggcggcgct caccagcctg acctcgatcg tcggacccct cctcttcacg
     6841 gcgatctatg cggcttctat aacaacgtgg aacgggtggg catggattgc aggcgctgcc
     6901 ctctacttgc tctgcctgcc ggcgctgcgt cgcgggcttt ggagcggcgc agggcaacga
     6961 gccgatcgct ga
//
