LOCUS       sequence-463764-        7673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BEVA Golden Gate cloning vector; BEVA2.0 Broad host range plasmid 
            with stability. L1-RK2-Tc-par-ELT4; GG assembly from pOGG010, 
            pOGG012, pOGG014, pOGG042, and pOGG004, TcR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7673)
  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 7673)
  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: pNDGG003
FEATURES             Location/Qualifiers
     source          1..7673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            140..248
                     /note="incP origin of transfer"
     CDS             complement(309..1457)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      1850..2381
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4903..4989)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    5077..5098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5113..5118,5119..5136,5137..5143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5151..5167
                     /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     5156..5178
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5175..5191
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5175..5191
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5187..5510
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    5204..5260
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5261..5278)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5261..5277)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5270..5292)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5486..5505)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      5615..5709
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5718..6368)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6474..7673
                     /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 ggccggccgc gatgcaggtg gctgctgaac ccccagccgg aactgacccc
      301 acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg
      361 ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg
      421 gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg
      481 tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc
      541 catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg
      601 acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac
      661 accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc
      721 gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg
      781 caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc
      841 aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg
      901 atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt
      961 ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc
     1021 gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc
     1081 gtgtgtttca gcaacgcggc ctgcttggct tcgctgacct gttttgccag gtcctcgccg
     1141 gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc
     1201 aaacctgccg cctcctgttc gagccgacgc gaacgctcca cggcggccga tggcgcgggc
     1261 agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggact
     1321 atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg
     1381 gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg
     1441 tcaaacgtcc gattcattca ccctccttgc gggattgccc cggaattaat tccccggatc
     1501 gatccgtcga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     1561 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     1621 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     1681 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     1741 ggggtcagca ccgtttctgc ggactggctt tctacgtggc tgccattttt ggggtgaggc
     1801 cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta gcgggccggg
     1861 agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca cagggcgcag
     1921 ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa gacaggttag
     1981 cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc tgtggacagc
     2041 ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc aagtgtcaag
     2101 gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg cagggcactt
     2161 atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc gttttcgccg
     2221 atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc ctcatctgtc
     2281 aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca tctgtcagtg
     2341 agggccaagt tttccgcgag gtatccacaa cgccggcggc cctacatggc tctgctgtag
     2401 tgagtgggtt gcgctccggc agcggtcctg atcccccgca gaaaaaaagg atctcaagaa
     2461 gatcctttga tcttttctac attactgcat gagcttgtgg aagtgtgctg agcataaatg
     2521 attctcctag ctgttctttg ggtaagtacg ccatcaggac gttgtgagtg gcgcgatttt
     2581 tagcggctga aatcagccct tgagcctgtc ggcaagtcgc gtcatgaggt ccatgcgctc
     2641 atgcaggatc gccacgacca acgcgggttc gcccgcacgc ggcaggcaaa aaacgtagtg
     2701 gtgttcgcag cgggccatcc gcagcgcggg aaagagttcg ctcatgtcct taaacgggcc
     2761 ttcgccggcg gcaagcctgg ctatgccctg ttccagctta gcgatatagc ggcgcacctg
     2821 cgccgcgccc cactcccggc gcgtgtagcg gatgatgccg cgtagatcgg cttcggcctc
     2881 agccgtgagg atgtaggccg tcaagcgcga tccccgctga gttcttcatc aagaatttcg
     2941 ccgacgctct tggtggacac cttgccggca agcccatcgt tgatgcggtt ccccagcatg
     3001 gttttcagtt cctgccatgc ctgatcggca tcagcgtcac cggggaacag acgttcgagg
     3061 gcgtattgct taatggtctt gccctgcaag gcggccaggg ctttcaggct ctggtgctgc
     3121 tggtccgtca tgtcgattgt caggcggctc attggataac ctccataaaa tacacgtaac
     3181 cacattagca catatgtggg cgtgaggcta cagcgcgagg cgcattaagg tcgggaaaat
     3241 gcgctaggcg catttaaatt gcgtattgct gtaatgcgcc atgccggcta gactaggccc
     3301 aaatgggtat acccaatttg accaaggggg acgcgatgag ggcggccaag cactaccgac
     3361 aacttctatc catcgacttc aacatcgagg cgctggcctt cgtgcctgga cccgacggca
     3421 cacgcggccg gcgcatccac gtcctggggc gcgaggtccg cgaccggccc ggcctggtcg
     3481 agtacctttc gccggcgttc ggctcgcggg tggcgctgga cggctactgc aaggccaatt
     3541 tcgatgcagt gctgcacctg gcgtaccccg atcatcagca atggggccac gcatgaagcg
     3601 ccgaagctac gccatgctgc gcgccgctgc cgcgctggcc gtcctggtcg ttgcctcgcc
     3661 ggcatgggcc gagctgcgcg gcgaggtcgt gcgcatcatc gacggcgaca ccatcgacgt
     3721 gctggtagac aagcagccgg tgcgcgtgcg cctggtggac attgacgcgc cggaaaagcg
     3781 gcaagccttc ggcgaacgtg cgcgccaggc gctggccggc atggtgttcc gccggcacgt
     3841 cctggtcgac gagaaggaca ccgaccgtta cggccgcacg ctgggcaccg tctgggtcaa
     3901 catggagctg gccagccggc cgccgcagcc gcgcaacgtc aacgccgcga tggttcacca
     3961 gggcatggcg tgggcctatc gcttccacgg ccgcgcggcc gaccctgaaa tgctgcggct
     4021 cgaacaggag gcgcgaggca agcgcgtcgg cctctggtcc gatccgcacg ccgtcgagcc
     4081 gtggaaatgg cgacgcgaga gcaacaaccg gagggacgaa ggttgaaggt cgcccgcatc
     4141 tacctgcgcg ccagtacgga cgagcagaat cttgaacgcc aggagagcct tgtagcggcc
     4201 acgcgggccg ccgggtacta cgtcgccggc atctaccgcg agaaggcgtc cggcgcacgc
     4261 gccgaccggc ccgagctgct gcgcatgatc gcggacctgc aacctggtga agtcgtcgtt
     4321 gcggagaaga tcgaccgcat cagccgcttg ccgttggccg aggccgagcg cctggttgcg
     4381 tcgatccggg ccaaaggggc caagctggcc gtgcctggcg tggtggacct gtcggagctg
     4441 gccgccgagg cgaacggagt ggcgaaaatc gttctggaat ccgtccagga catgcttttg
     4501 aagctcgcct tgcagatggc ccgcgacgac tacgaggatc ggcgcgagcg tcaacgtcag
     4561 ggtgtccagt tggcgaaggc cgccggccgc tacaccggcc gcaaacgtga cgccggcatg
     4621 cacgaccgca tcatcacgct tcgctccggc ggatcgagca ttgccaagac ggccaagctg
     4681 gtcggatgca gcccgagcca ggtcaaacga gtgtgggcgg cctggaacgc gcagcagcaa
     4741 aaataaagcc gggcagtgcc cggcttttct caccttttcg cgtcccgcag ggccgctgcg
     4801 agcgccctac ctagatcctc gctttccccc tcggtgtagt ccggccaggg cacgaagggc
     4861 gcggatgcga acctgttgag caggtcagat ctagggcggc ggatttgtcc tactcaggag
     4921 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     4981 ttttatttga tgcctgccga attcggatcc ggagtgagac cgcagctggc acgacaggtt
     5041 tgccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     5101 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5161 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
     5221 ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
     5281 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     5341 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     5401 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5461 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5521 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5581 cttacagaca agctgtgacg gtctcacgct cttggactcc tgttgataga tccagtaatg
     5641 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
     5701 ggtgagaatc caggcaatca atcgtcaccc tttctcggtc cttcaacgtt cctgacaacg
     5761 agcctccttt tcgccaatcc atcgacaatc accgcgagtc cctgctcgaa cgctgcgtcc
     5821 ggaccggctt cgtcgaaggc gtctatcgcg gcccgcaaca gcggcgagag cggagcctgt
     5881 tcaacggtgc cgccgcgctc gccggcatcg ctgtcgccgg cctgctcctc aagcacggcc
     5941 ccaacagtga agtagctgat tgtcatcagc gcattgacgg cgtccccggc cgaaaaaccc
     6001 gcctcgcaga ggaagcgaag ctgcgcgtcg gccgtttcca tctgcggtgc gcccggtcgc
     6061 gtgccggcat ggatgcgcgc gccatcgcgg taggcgagca gcgcctgcct gaagctgcgg
     6121 gcattcccga tcagaaatga gcgccagtcg tcgtcggctc tcggcaccga atgcgtatga
     6181 ttctccgcca gcatggcttc ggccagtgcg tcgagcagcg cccgcttgtt cctgaagtgc
     6241 cagtaaagcg ccggctgctg aacccccaac cgttccgcca gtttgcgtgt cgtcagaccg
     6301 tctacgccga cctcgttcaa caggtccagg gcggcacgga tcactgtatt cggctgcaac
     6361 tttgtcatgc ttgacacttt atcactgata aacataatat gtccaccaac ttatcagtga
     6421 taaagaatcc gcgcgttcaa tcggaccagc ggaggctggt ccggaggcca gacgtgaaac
     6481 ccaacatacc cctgatcgta attctgagca ctgtcgcgct cgacgctgtc ggcatcggcc
     6541 tgattatgcc ggtgctgccg ggcctcctgc gcgatctggt tcactcgaac gacgtcaccg
     6601 cccactatgg cattctgctg gcgctgtatg cgttggtgca atttgcctgc gcacctgtgc
     6661 tgggcgcgct gtcggatcgt ttcgggcggc ggccaatctt gctcgtgtcg ctggccggcg
     6721 ccactgtcga ctacgccatc atggcgacag cgcctttcct ttgggttctc tatatcgggc
     6781 ggatcgtggc cggcatcacc ggggcgactg gggcggtagc cggcgcttat attgccgata
     6841 tcactgatgg cgatgagcgc gcgcggcact tcggcttcat gagcgcctgt ttcgggttcg
     6901 ggatggtcgc gggacctgtg ctcggtgggc tgatgggcgg tttctccccc cacgctccgt
     6961 tcttcgccgc ggcagccttg aacggcctca atttcctgac gggctgtttc cttttgccgg
     7021 agtcgcacaa aggcgaacgc cggccgttac gccgggaggc tctcaacccg ctcgcttcgt
     7081 tccggtgggc ccggggcatg accgtcgtcg ccgccctgat ggcggtgttc ttcatcatgc
     7141 aacttgtcgg acaggtgccg gccgcgcttt gggtcatttt cggcgaggat cgctttcact
     7201 gggacgcgac cacgatcggc atttcgcttg ccgcatttgg cattctgcat tcactcgccc
     7261 aggcaatgat caccggccct gtagccgccc ggctcggcga aaggcgggca ctcatgctcg
     7321 gaatgattgc cgacggcaca ggctacatcc tgcttgcctt cgcgacacgg ggatggatgg
     7381 cgttcccgat catggtcctg cttgcttcgg gtggcatcgg aatgccggcg ctgcaagcaa
     7441 tgttgtccag gcaggtggat gaggaacgtc aggggcagct gcaaggctca ctggcggcgc
     7501 tcaccagcct gacctcgatc gtcggacccc tcctcttcac ggcgatctat gcggcttcta
     7561 taacaacgtg gaacgggtgg gcatggattg caggcgctgc cctctacttg ctctgcctgc
     7621 cggcgctgcg tcgcgggctt tggagcggcg cagggcaacg agccgatcgc tga
//
