LOCUS       sequence-223395-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector construction module. RK2 (IncP) low-copy number origin of 
            replication for construction of Golden Gate cloning vectors .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5040)
  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 5040)
  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: pOGG010
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     oriT            539..647
                     /note="incP origin of transfer"
     CDS             complement(708..1856)
                     /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      2249..2780
                     /label=oriV
                     /note="incP origin of replication"
     primer_bind     complement(3030..3047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3140..3728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3220..3239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(4913..5017)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattgagtg aaggccgtca
      361 aggcctaggc gcgccatgag ctccactctg tgcgtctcaa ctacttttcc gctgcataac
      421 cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata
      481 caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg
      541 gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt
      601 cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggccgcg
      661 atgcaggtgg ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa
      721 tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg
      781 caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg
      841 aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac
      901 atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg
      961 tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc
     1021 ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg
     1081 cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc
     1141 gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag
     1201 gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt
     1261 tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg
     1321 tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac
     1381 agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata
     1441 tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc
     1501 tgcttggctt cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc
     1561 gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttcg
     1621 agccgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc
     1681 acgctgtcgc gctcgatctt ggccgtagct tgctggacta tcgagccgac ggactggaag
     1741 gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa
     1801 aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac
     1861 cctccttgcg ggattgcccc ggaattaatt ccccggatcg atccgtcgat cttgatcccc
     1921 tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa
     1981 ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc
     2041 agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt
     2101 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     2161 gactggcttt ctacgtggct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca
     2221 gcccctgggg ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca
     2281 ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt
     2341 ataaatattg gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg
     2401 aaacccttgc aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc
     2461 gcccctcatc tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca
     2521 gtagtcgcgc ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc
     2581 atctgtggga aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc
     2641 cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg
     2701 gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg
     2761 tatccacaac gccggcggcc ctacatggct ctgctgtagt gagtgggttg cgctccggca
     2821 gcggtcctga tcccccgcag aaaaaaagga tctcaagaag atcctttgat cttttctaca
     2881 ttactgagac gcacgaagtg ggtacctctt aattaactgg cctcatgggc cttcacttca
     2941 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg tcatagctgt
     3001 ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3061 cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3901 attatttacc caccactttg gtaatttcgc ctttcacgta atgcacaaat tcttccagct
     3961 gatccgcacg gctggccaga cgatcttctt cctggcccag atacgcctga cgcgcttcca
     4021 gaatcaccgg ctgatactgc gccggcagac gttccatggc ccaatccgcc gccacatctt
     4081 tcggcgcaat tttgccggtc accgcgctat accaaatacg gctcagggtc agcaccacgt
     4141 tgcgttcatc acccgcccaa tccggcgggc tgttccacag ggtcagggtt tcgttcagcg
     4201 cttcaaacag gtcctgttcc ggcaccggat caaacagttc ttccgctgcc ggaccaacca
     4261 gcgccacgct atgttcacgc gctttggtca gcagaatggc cagatcgata tcaatggtcg
     4321 ccggttcaaa aatgccggcc agaatatcgt tacgctgcca ttcgccaaac tgcagctcac
     4381 gtttcgccgg ataacgccac ggaataatgt catcatgcac cacgatggtc acttccaccg
     4441 cacgcagaat ttcgctttca cccgggctcg cgctggtttc cagcagatcg ttaatcagcg
     4501 cacgacgggt ggtttcatcc agacgcacgg tcacggtcac cagcagatca atatcgctat
     4561 gcggtttcag gccgccatcc accgcgctgc catacagatg cacggccagc agggtcggtt
     4621 ccagatgacg ttcaatcacg cccaccactt cagacagctg ggtgctcact tccgcaatca
     4681 ccgcttcacg cataatgttc agtttggttt tcggacggct gccctgctgg gtcacatcgt
     4741 tgctgctcca cagcatcagg caacggccca ccgcataacg cgcctgctgt ttgctcgcac
     4801 gcggcatgct ctgcacgcca aaaaagcaat catagcacgc cataaacacc gccaccgcac
     4861 cattaccgcc gctacgttcg gtcagggtac ggctccaatt acggctacgc atactcttcc
     4921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
