LOCUS       sequence-224356-        9574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 golden gate cloning vector for medium copy number 
            free-living expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9574)
  AUTHORS   Geddes BA, Mendoza-Suarez MA, Poole PS
  TITLE     A Bacterial Expression Vector Archive (BEVA) for Flexible Modular 
            Assembly of Golden Gate-Compatible Vectors.
  JOURNAL   Front Microbiol. 2019 Jan 14;9:3345. doi: 10.3389/fmicb.2018.03345. 
            eCollection 2018.
  PUBMED    30692983
REFERENCE   2  (bases 1 to 9574)
  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: "Front 
            Microbiol."; date: "2019-01-14"; volume: "9:3345. doi"; pages: " 
            10.3389/fmicb.2018.03345. eCollection 2018"
COMMENT     Sequence Label: pOGG216
FEATURES             Location/Qualifiers
     source          1..9574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1532..1541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      5627..5721
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5730..6380)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             6486..7685
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     oriT            7825..7933
                     /note="incP origin of transfer"
     CDS             complement(7944..8606)
                     /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      8607..9378
                     /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(9483..9569)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcctgcctt gtcttctggt cgactcgacg gagggtaccg cacggtctgc caatccgacg
       61 gaggtttatg aattttccac cttttccaca agctcaacta gtattaacga tgtggattta
      121 gcaaaaaaaa cctgtaaccc taaatgtaaa ataacgggta agcctgccaa ccatgttatg
      181 gcagattaag cgtctttttg aagggcaccg catctttcgc gttgccgtaa atgtatccgt
      241 ttataaggac agcccgaatg acggtctgcg caaaaaaaca cgttcatctc actcgcgatg
      301 ctgcggagca gttactggct gatattgatc gacgccttga tcagttattg cccgtggagg
      361 gagaacggga tgttgtgggt gccgcgatgc gtgaaggtgc gctggcaccg ggaaaacgta
      421 ttcgccccat gttgctgttg ctgaccgccc gcgatctggg ttgcgctgtc agccatgacg
      481 gattactgga tttggcctgt gcggtggaaa tggtccacgc ggcttcgctg atccttgacg
      541 atatgccctg catggacgat gcgaagctgc ggcgcggacg ccctaccatt cattctcatt
      601 acggagagca tgtggcaata ctggcggcgg ttgccttgct gagtaaagcc tttggcgtaa
      661 ttgccgatgc agatggcctc acgccgctgg caaaaaatcg ggcggtttct gaactgtcaa
      721 acgccatcgg catgcaagga ttggttcagg gtcagttcaa ggatctgtct gaaggggata
      781 agccgcgcag cgctgaagct attttgatga cgaatcactt taaaaccagc acgctgtttt
      841 gtgcctccat gcagatggcc tcgattgttg cgaatgcctc cagcgaagcg cgtgattgcc
      901 tgcatcgttt ttcacttgat cttggtcagg catttcaact gctggacgat ttgaccgatg
      961 gcatgaccga caccggtaag gatagcaatc aggacgccgg taaatcgacg ctggtcaatc
     1021 tgttaggccc gagggcggtt gaagaacgtc tgagacaaca tcttcagctt gccagtgagc
     1081 atctctctgc ggcctgccaa cacgggcacg ccactcaaca ttttattcag gcctggtttg
     1141 acaaaaaact cgctgccgtc agttaaggat gctgcatgag cgcacatgcc ctgcccaagg
     1201 cagatctgac cgccaccagc ctgatcgtat cgggcggcat catcgccgct tggctggccc
     1261 tgcatgtgca tgcgctgtgg tttctggacg cagcggcgca tcccatcctg gcgatcgcaa
     1321 atttcctggg gctgacctgg ctgtcggtcg gattgttcat catcgcgcat gacgcgatgc
     1381 acgggtcggt ggtgccgggg cgtccgcgcg ccaatgcggc gatgggccag cttgtcctgt
     1441 ggctgtatgc cggattttcg tggcgcaaga tgatcgtcaa gcacatggcc catcaccgcc
     1501 atgccggaac cgacgacgac cccgatttcg accatggcgg cccggtccgc tggtacgccc
     1561 gcttcatcgg cacctatttc ggctggcgcg aggggctgct tctgcccgtc atcgtgacgg
     1621 tctatgcgct gatccttggg gatcgctgga tgtacgtggt attctggccg ctgccgtcga
     1681 tcctggcgtc gatccagctg ttcgtgttcg gcacctggct gccgcaccgc cccggccacg
     1741 acgcgttccc ggaccgccac aatgcgcggt cgtcgcggat cagcgacccc gtgtcgctgc
     1801 tgacctgttt tcactttggc ggttatcatc acgaacacca cctgcacccg acggtgccgt
     1861 ggtggcgcct gcccagcacc cgcaccaagg gggacaccgc atgagtgact taagtgggag
     1921 cggctatgca accgcattat gatctgattc tcgtgggggc tggactcgcg aatggcctta
     1981 tcgccctgcg acttcagcag cagcaacctg atatgcgtat tttgcttatc gacgccgcac
     2041 cccaggcggg cgggaatcat acgtggtcat ttcaccacga tgatttgact gagagccaac
     2101 atcgttggat agctccgctg gtggttcatc actggcccga ctatcaggta cgctttccca
     2161 cacgccgtcg taagctgaac agcggctact tttgtattac ttctcagcgt ttcgctgagg
     2221 ttttacagcg acagtttggc ccgcacttgt ggatggatac cgcggtcgca gaggttaatg
     2281 cggaatctgt tcggttgaaa aagggtcagg ttatcggtgc ccgcgcggtg attgacgggc
     2341 ggggttatgc ggcaaattca gcactgagcg tgggcttcca ggcgtttatt ggccaggaat
     2401 ggcgattgag ccacccgcat ggtttatcat ctcccattat catggatgcc acggtcgatc
     2461 agcaaaatgg ttatcgcttc gtgtacagcc tgccgctctc gccgaccaga ttgttaattg
     2521 aagatacgca ctatattgat aatgcgacat tagatcctga atgcgcgcgg caaaatattt
     2581 gcgactatgc cgcgcaacag ggttggcagc ttcagacact gctgcgagaa gaacagggcg
     2641 ccttacccat tactctgtcg ggcaatgccg acgcattctg gcagcagcgc cccctggcct
     2701 gtagtggatt acgtgccggt ctgttccatc ctaccaccgg ctattcactg ccgctggcgg
     2761 ttgccgtggc cgaccgcctg agtgcacttg atgtctttac gtcggcctca attcaccatg
     2821 ccattacgca ttttgcccgc gagcgctggc agcagcaggg ctttttccgc atgctgaatc
     2881 gcatgctgtt tttagccgga cccgccgatt cacgctggcg ggttatgcag cgtttttatg
     2941 gtttacctga agatttaatt gcccgttttt atgcgggaaa actcacgctg accgatcggc
     3001 tacgtattct gagcggcaag ccgcctgttc cggtattagc agcattgcaa gccattatga
     3061 cgactcatcg ttaaagagcg actacatgaa accaactacg gtaattggtg caggcttcgg
     3121 tggcctggca ctggcaattc gtctacaagc tgcggggatc cccgtcttac tgcttgaaca
     3181 acgtgataaa cccggcggtc gggcttatgt ctacgaggat caggggttta cctttgatgc
     3241 aggcccgacg gttatcaccg atcccagtgc cattgaagaa ctgtttgcac tggcaggaaa
     3301 acagttaaaa gagtatgtcg aactgctgcc ggttacgccg ttttaccgcc tgtgttggga
     3361 gtcagggaag gtctttaatt acgataacga tcaaacccgg ctcgaagcgc agattcagca
     3421 gtttaatccc cgcgatgtcg aaggttatcg tcagtttctg gactattcac gcgcggtgtt
     3481 taaagaaggc tatctaaagc tcggtactgt ccctttttta tcgttcagag acatgcttcg
     3541 cgccgcacct caactggcga aactgcaggc atggagaagc gtttacagta aggttgccag
     3601 ttacatcgaa gatgaacatc tgcgccaggc gttttctttc cactcgctgt tggtgggcgg
     3661 caatcccttc gccacctcat ccatttatac gttgatacac gcgctggagc gtgagtgggg
     3721 cgtctggttt ccgcgtggcg gcaccggcgc attagttcag gggatgataa agctgtttca
     3781 ggatctgggt ggcgaagtcg tgttaaacgc cagagtcagc catatggaaa cgacaggaaa
     3841 caagattgaa gccgtgcatt tagaggacgg tcgcaggttc ctgacgcaag ccgtcgcgtc
     3901 aaatgcagat gtggttcata cctatcgcga cctgttaagc cagcaccctg ccgcggttaa
     3961 gcagtccaac aaactgcaga ctaagcgcat gagtaactct ctgtttgtgc tctattttgg
     4021 tttgaatcac catcatgatc agctcgcgca tcacacggtt tgtttcggcc cgcgttaccg
     4081 cgagctgatt gacgaaattt ttaatcatga tggcctcgca gaggacttct cactttatct
     4141 gcacgcgccc tgtgtcacgg attcgtcact ggcgcctgaa ggttgcggca gttactatgt
     4201 gttggcgccg gtgccgcatt taggcaccgc gaacctcgac tggacggttg aggggccaaa
     4261 actacgcgac cgtatttttg cgtaccttga gcagcattac atgcctggct tacggagtca
     4321 gctggtcacg caccggatgt ttacgccgtt tgattttcgc gaccagctta atgcctatca
     4381 tggctcagcc ttttctgtgg agcccgttct tacccagagc gcctggtttc ggccgcataa
     4441 ccgcgataaa accattacta atctctacct ggtcggcgca ggcacgcatc ccggcgcagg
     4501 cattcctggc gtcatcggct cggcaaaagc gacagcaggt ttgatgctgg aggatctgat
     4561 ttgaataatc cgtcgttact caatcatgcg gtcgaaacga tggcagttgg ctcgaaaagt
     4621 tttgcgacag cctcaaagtt atttgatgca aaaacccggc gcagcgtact gatgctctac
     4681 gcctggtgcc gccattgtga cgatgttatt gacgatcaga cgctgggctt tcaggcccgg
     4741 cagcctgcct tacaaacgcc cgaacaacgt ctgatgcaac ttgagatgaa aacgcgccag
     4801 gcctatgcag gatcgcagat gcacgaaccg gcgtttgcgg cttttcagga agtggctatg
     4861 gctcatgata tcgccccggc ttacgcgttt gatcatctgg aaggcttcgc catggatgta
     4921 cgcgaagcgc aatacagcca actggatgat acgctgcgct attgctatca cgttgcaggc
     4981 gttgtcggct tgatgatggc gcaaatcatg ggcgtgcggg ataacgccac gctggaccgc
     5041 gcctgtgacc ttgggctggc atttcagttg accaatattg ctcgcgatat tgtggacgat
     5101 gcgcatgcgg gccgctgtta tctgccggca agctggctgg agcatgaagg tctgaacaaa
     5161 gagaattatg cggcacctga aaaccgtcag gcgctgagcc gtatcgcccg tcgtttggtg
     5221 caggaagcag aaccttacta tttgtctgcc acagccggcc tggcagggtt gcccctgcgt
     5281 tccgcctggg caatcgctac ggcgaagcag gtttaccgga aaataggtgt caaagttgaa
     5341 caggccggtc agcaagcctg ggatcagcgg cagtcaacga ccacgcccga aaaattaacg
     5401 ctgctgctgg ccgcctctgg tcaggccctt acttcccgga tgcgggctca tcctccccgc
     5461 cctgcgcatc tctggcagcg cccgctctag cgccatgtct ttcccggagc gtcgcctgaa
     5521 gttttgacag gggcggcgca tagaggaagc caaaagaaac acaaccttct ttgcccctga
     5581 cggcgtgatg catacgctgg cgattagcac gaagacaagg gacttggact cctgttgata
     5641 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     5701 gcgtttttta ttggtgagaa tccaggcaat caatcgtcac cctttctcgg tccttcaacg
     5761 ttcctgacaa cgagcctcct tttcgccaat ccatcgacaa tcaccgcgag tccctgctcg
     5821 aacgctgcgt ccggaccggc ttcgtcgaag gcgtctatcg cggcccgcaa cagcggcgag
     5881 agcggagcct gttcaacggt gccgccgcgc tcgccggcat cgctgtcgcc ggcctgctcc
     5941 tcaagcacgg ccccaacagt gaagtagctg attgtcatca gcgcattgac ggcgtccccg
     6001 gccgaaaaac ccgcctcgca gaggaagcga agctgcgcgt cggccgtttc catctgcggt
     6061 gcgcccggtc gcgtgccggc atggatgcgc gcgccatcgc ggtaggcgag cagcgcctgc
     6121 ctgaagctgc gggcattccc gatcagaaat gagcgccagt cgtcgtcggc tctcggcacc
     6181 gaatgcgtat gattctccgc cagcatggct tcggccagtg cgtcgagcag cgcccgcttg
     6241 ttcctgaagt gccagtaaag cgccggctgc tgaaccccca accgttccgc cagtttgcgt
     6301 gtcgtcagac cgtctacgcc gacctcgttc aacaggtcca gggcggcacg gatcactgta
     6361 ttcggctgca actttgtcat gcttgacact ttatcactga taaacataat atgtccacca
     6421 acttatcagt gataaagaat ccgcgcgttc aatcggacca gcggaggctg gtccggaggc
     6481 cagacgtgaa acccaacata cccctgatcg taattctgag cactgtcgcg ctcgacgctg
     6541 tcggcatcgg cctgattatg ccggtgctgc cgggcctcct gcgcgatctg gttcactcga
     6601 acgacgtcac cgcccactat ggcattctgc tggcgctgta tgcgttggtg caatttgcct
     6661 gcgcacctgt gctgggcgcg ctgtcggatc gtttcgggcg gcggccaatc ttgctcgtgt
     6721 cgctggccgg cgccactgtc gactacgcca tcatggcgac agcgcctttc ctttgggttc
     6781 tctatatcgg gcggatcgtg gccggcatca ccggggcgac tggggcggta gccggcgctt
     6841 atattgccga tatcactgat ggcgatgagc gcgcgcggca cttcggcttc atgagcgcct
     6901 gtttcgggtt cgggatggtc gcgggacctg tgctcggtgg gctgatgggc ggtttctccc
     6961 cccacgctcc gttcttcgcc gcggcagcct tgaacggcct caatttcctg acgggctgtt
     7021 tccttttgcc ggagtcgcac aaaggcgaac gccggccgtt acgccgggag gctctcaacc
     7081 cgctcgcttc gttccggtgg gcccggggca tgaccgtcgt cgccgccctg atggcggtgt
     7141 tcttcatcat gcaacttgtc ggacaggtgc cggccgcgct ttgggtcatt ttcggcgagg
     7201 atcgctttca ctgggacgcg accacgatcg gcatttcgct tgccgcattt ggcattctgc
     7261 attcactcgc ccaggcaatg atcaccggcc ctgtagccgc ccggctcggc gaaaggcggg
     7321 cactcatgct cggaatgatt gccgacggca caggctacat cctgcttgcc ttcgcgacac
     7381 ggggatggat ggcgttcccg atcatggtcc tgcttgcttc gggtggcatc ggaatgccgg
     7441 cgctgcaagc aatgttgtcc aggcaggtgg atgaggaacg tcaggggcag ctgcaaggct
     7501 cactggcggc gctcaccagc ctgacctcga tcgtcggacc cctcctcttc acggcgatct
     7561 atgcggcttc tataacaacg tggaacgggt gggcatggat tgcaggcgct gccctctact
     7621 tgctctgcct gccggcgctg cgtcgcgggc tttggagcgg cgcagggcaa cgagccgatc
     7681 gctgaactac ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
     7741 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
     7801 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     7861 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     7921 gcgaggctgg ccgtaggccg gccctaccgg cgcggcagcg ttacccgtgt cggcggctcc
     7981 aacggctcgc catcgtccag aaaacacggc tcatcgggca tcggcaggcg ctgctgcccg
     8041 cgccgttccc attcctccgt ttcggtcaag gctggcaggt ctggttccat gcccggaatg
     8101 ccgggctggc tgggcggctc ctcgccgggg ccggtcggta gttgctgctc gcccggatac
     8161 agggtcggga tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg gtcgtcgtga
     8221 tcaaccacca cggcggcact gaacaccgac aggcgcaact ggtcgcgggg ctggccccac
     8281 gccacgcggt cattgaccac gtaggccgac acggtgccgg ggccgttgag cttcacgacg
     8341 gagatccagc gctcggccac caagtccttg actgcgtatt ggaccgtccg caaagaacgt
     8401 ccgatgagct tggaaagagt cttttggctg accaccacgg cgttctggtg gcccatctgc
     8461 gcgacgaggt gatgcagcag cattgccgcc gtgggtttcc tcgcaataag cccggcccac
     8521 gcctcatgcg ctttgcgttc cgtttggacc cagtgaccgg gcttgttctt ggcttgaatg
     8581 ccgatttctc tggactgcgt ggccatgctt atctccatgc ggtaggggtg ccgcacggtt
     8641 gcggcaccat gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt atgcgttgcg
     8701 taaaagtggc agtcaattac agattttctt taacctacgc aatgagctat tgcggggggt
     8761 gccgcaatga gctgttgcgt accccccttt tttaagttgt tgatttttaa gtctttcgca
     8821 tttcgcccta tatctagttc tttggtgccc aaagaagggc acccctgcgg ggttccccca
     8881 cgccttcggc gcggctcccc ctccggcaaa aagtggcccc tccggggctt gttgatcgac
     8941 tgcgcggcct tcggccttgc ccaaggtggc gctgccccct tggaaccccc gcactcgccg
     9001 ccgtgaggct cggggggcag gcgggcgggc ttcgcccttc gactgccccc actcgcatag
     9061 gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc gcgagccttg
     9121 acccgccttc ctcttggtgt ccaaccggca agcgaagcgc gcaggccgca ggccggaggc
     9181 ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc gcagttggag
     9241 ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg tcaagctcgt
     9301 gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc
     9361 gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg caagggagcg
     9421 cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggggttact ctagggcggc
     9481 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     9541 tctttcgact gagcctttcg ttttatttga tgcc
//
