LOCUS       sequence-472146-        8471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector part for BEVA; BEVA2.0 Position 3 RSF1010 broad host range 
            origin of replication with oriT, CmR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8471)
  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 8471)
  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: pNDGG005
FEATURES             Location/Qualifiers
     source          1..8471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        254..272
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     254..271
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     oriT            465..573
                     /note="incP origin of transfer"
     rep_origin      1260..1654
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            1996..2082
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             3321..4292
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLASFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEETGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             4803..5642
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             5629..6480
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDALCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     primer_bind     complement(6714..6733)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6715..6733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(6844..7503)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7418..7437
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7504..7606)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      7707..8295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8196..8215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8449..8466
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgaaaaatca ataatcagac aacaagatgt gcgaactcga tattttacac gactctcttt
       61 accaattctg ccccgaatta cacttaaaac gactcaacag cttaacgttg gcttgccacg
      121 cattacttga ctgtaaaact ctcactctta ccgaacttgg ccgtaacctg ccaaccaaag
      181 cgagaacaaa acataacatc aaacgaatcg accgattgtt aggtaatcgt cacctgcagg
      241 aaggtttaaa cgcatttagg tgacactata gaagtgtgta tcgctcgagg gatccgaatt
      301 cgaagtcttg gtacggagcg tctcaactac ttttccgctg cataaccctg cttcggggtc
      361 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
      421 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
      481 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
      541 ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccgcgatca agcaggtgcg
      601 acagacgtca tactagatat caagcgactt ctcctatccc ctgggaacac atcaatctca
      661 ccggagaata tcgctggcca aagccttagc gtaggattcc gccccttccc gcaaacgacc
      721 ccaaacagga aacgcagctg aaacgggaag ctcaacaccc actgacgcat gggttgttca
      781 ggcagtactt catcaaccag caaggcggca ctttcggcca tccgccgcgc cccacagctc
      841 gggcagaaac cgcgacgctt acagctgaaa gcgaccaggt gctcggcgtg gcaagactcg
      901 cagcgaaccc gtagaaagcc atgctccagc cgcccgcatt ggagaaattc ttcaaattcc
      961 cgttgcacat agcccggcaa ttcctttccc tgctctgcca taagcgcagc gaatgccggg
     1021 taatactcgt caacgatctg atagagaagg gtttgctcgg gtcggtggct ctggtaacga
     1081 ccagtatccc gatcccggct ggccgtcctg gccgccacat gaggcatgtt ccgcgtcctt
     1141 gcaatactgt gtttacatac agtctatcgc ttagcggaaa gttcttttac cctcagccga
     1201 aatgcctgcc gttgctagac attgccagcc agtgcccgtc actcccgtac taactgtcac
     1261 gaacccctgc aataactgtc acgcccccct gcaataactg tcacgaaccc ctgcaataac
     1321 tgtcacgccc ccaaacctgc aaacccagca ggggcggggg ctggcggggt gttggaaaaa
     1381 tccatccatg attatctaag aataatccac taggcgcggt tatcagcgcc cttgtggggc
     1441 gctgctgccc ttgcccaata tgcccggcca gaggccgggt agctggtcta ttcgctgcgc
     1501 taggctacac accgccccac cgctgcgcgg cagggggaaa ggcgggcaaa gcccgctaaa
     1561 ccccacacca aaccccgcag aaatacgctg gagcgctttt agccgcttta gcggcctttc
     1621 cccctacccg aagggtgggg gcgcgtgtgc agccccgcag ggcctgtctc ggtcgatcat
     1681 tcagcccggc tcatccttct ggcgtggcgg cagaccgaac aaggcgcggt cgtggtcgcg
     1741 ttcaaggtac gcatccattg ccgccatgag ccgatcctcc ggccactcgc tgctgttcac
     1801 cttggccaaa atcatggccc ccaccagcac cctgcgcctt gtttcgttct tgcgctcttg
     1861 ctgctgttcc cttgcccgca cccgctgaat ttcggcattg attcgcgctc gttgttcttc
     1921 gagcttggcc agccgatccg ccgccttgtt gctcccctta accatcttga caccccattg
     1981 ttaatgtgct gtctcgtagg ctatcatgga ggcacagcgg cggcaatccc gaccctactt
     2041 tgtaggggag gggcacttac cggtttctct tcgagaaact ggcctaacgg ccacccttcg
     2101 ggcggtgcgc tctccgaggg ccattgcatg gagccgaaaa gcaaaagcaa cagcgaggca
     2161 gcatggcgat ttatcacctt acggcgaaaa ccggcagcag gtcgggcggc caatcggcca
     2221 gggccaaggc cgactacatc cagcgcgaag gcaagtatgc ccgcgacatg gatgaagtct
     2281 tgcacgccga atccgggcac atgccggagt tcgtcgagcg gcccgccgac tactgggatg
     2341 ctgccgacct gtatgaacgc gccaatgggc ggctgttcaa ggaggtcgaa tttgccctgc
     2401 cggtcgagct gaccctcgac cagcagaagg cgctggcgtc cgagttcgcc cagcacctga
     2461 ccggtgccga gcgcctgccg tatacgctgg ccatccatgc cggtggcggc gagaacccgc
     2521 actgccacct gatgatctcc gagcggatca atgacggcat cgagcggccc gccgctcagt
     2581 ggttcaagcg gtacaacggc aagaccccgg agaagggcgg ggcacggaag accgaagcgc
     2641 tcaagcccaa ggcatggctt gagcagaccc gcgaggcatg ggccgaccat gccaaccggg
     2701 cattagagcg ggctggccac gacgcccgca ttgaccacag aacacttgag gcgcagggca
     2761 tcgagcgcct gcccggtgtt cacctggggc cgaacgtggt ggagatggaa ggccggggca
     2821 tccgcaccga ccgggcagac gtggccctga acatcgacac cgccaacacc cagatcatcg
     2881 acttacagga ataccgggag gcaatagacc atgaacgcaa tcgacagagt gaagaaatcc
     2941 agaggcatca acgagttagc ggagcagatc gaaccgctgg cccagagcat ggcgacactg
     3001 gccgacgaag cccggcaggt catgagccag acccagcagg ccagcgaggc gcaggcggcg
     3061 gagtggctga aagcccagcg ccagacaggg gcggcatggg tggagctggc caaagagttg
     3121 cgggaggtag ccgccgaggt gagcagcgcc gcgcagagcg cccggagcgc gtcgcggggg
     3181 tggcactgga agctatggct aaccgtgatg ctggcttcca tgatgcctac ggtggtgctg
     3241 ctgatcgcat cgttgctctt gctcgacctg acgccactga caaccgagga cggctcgatc
     3301 tggctgcgct tggtggcccg atgaagaacg acaggacttt gcaggccata ggccgacagc
     3361 tcaaggccat gggctgtgag cgcttcgata tcggcgtcag ggacgccacc accggccaga
     3421 tgatgaaccg ggaatggtca gccgccgaag tgctccagaa cacgccatgg ctcaagcgga
     3481 tgaatgccca gggcaatgac gtgtatatca ggcccgccga gcaggagcgg catggtctgg
     3541 tgctggtgga cgacctcagc gagtttgacc tggatgacat gaaagccgag ggccgggagc
     3601 ctgccctggt agtggaaacc agcccgaaga actatcaggc atgggtcaag gtggccgacg
     3661 ccgcaggcgg tgaacttcgg gggcagattg cccggacgct ggccagcgag tacgacgccg
     3721 acccggccag cgccgacagc cgccactatg gccgcttggc gagcttcacc aaccgcaagg
     3781 acaagcacac cacccgcgcc ggttatcagc cgtgggtgct gctgcgtgaa tccaagggca
     3841 agaccgccac cgctggcccg gcgctggtgc agcaggctgg ccagcagatc gagcaggccc
     3901 agcggcagca ggagaaggcc cgcaggctgg ccagcctcga actgcccgag cggcagctta
     3961 gccgccaccg gcgcacggcg ctggacgagt accgcagcga gatggccggg ctggtcaagc
     4021 gcttcggtga tgacctcagc aagtgcgact ttatcgccgc gcagaagctg gccagccggg
     4081 gccgcagtgc cgaggaaacc ggcaaggcca tggccgaggc cagcccagcg ctggcagagc
     4141 gcaagcccgg ccacgaagcg gattacatcg agcgcaccgt cagcaaggtc atgggtctgc
     4201 ccagcgtcca gcttgcgcgg gccgagctgg cacgggcacc ggcaccccgc cagcgaggca
     4261 tggacagggg cgggccagat ttcagcatgt agtgcttgcg ttggtactca cgcctgttat
     4321 actatgagta ctcacgcaca gaagggggtt ttatggaata cgaaaaaagc gcttcagggt
     4381 cggtctacct gatcaaaagt gacaagggct attggttgcc cggtggcttt ggttatacgt
     4441 caaacaaggc cgaggctggc cgcttttcag tcgctgatat ggccagcctt aaccttgacg
     4501 gctgcacctt gtccttgttc cgcgaagaca agcctttcgg ccccggcaag tttctcggtg
     4561 actgatatga aagaccaaaa ggacaagcag accggcgacc tgctggccag ccctgacgct
     4621 gtacgccaag cgcgatatgc cgagcgcatg aaggccaaag ggatgcgtca gcgcaagttc
     4681 tggctgaccg acgacgaata cgaggcgctg cgcgagtgcc tggaagaact cagagcggcg
     4741 cagggcgggg gtagtgaccc cgccagcgcc taaccaccaa ctgcctgcaa aggaggcaat
     4801 caatggctac ccataagcct atcaatattc tggaggcgtt cgcagcagcg ccgccaccgc
     4861 tggactacgt tttgcccaac atggtggccg gtacggtcgg ggcgctggtg tcgcccggtg
     4921 gtgccggtaa atccatgctg gccctgcaac tggccgcaca gattgcaggc gggccggatc
     4981 tgctggaggt gggcgaactg cccaccggcc cggtgatcta cctgcccgcc gaagacccgc
     5041 ccaccgccat tcatcaccgc ctgcacgccc ttggggcgca cctcagcgcc gaggaacggc
     5101 aagccgtggc tgacggcctg ctgatccagc cgctgatcgg cagcctgccc aacatcatgg
     5161 ccccggagtg gttcgacggc ctcaagcgcg ccgccgaggg ccgccgcctg atggtgctgg
     5221 acacgctgcg ccggttccac atcgaggaag aaaacgccag cggccccatg gcccaggtca
     5281 tcggtcgcat ggaggccatc gccgccgata ccgggtgctc tatcgtgttc ctgcaccatg
     5341 ccagcaaggg cgcggccatg atgggcgcag gcgaccagca gcaggccagc cggggcagct
     5401 cggtactggt cgataacatc cgctggcagt cctacctgtc gagcatgacc agcgccgagg
     5461 ccgaggaatg gggtgtggac gacgaccagc gccggttctt cgtccgcttc ggtgtgagca
     5521 aggccaacta tggcgcaccg ttcgctgatc ggtggttcag gcggcatgac ggcggggtgc
     5581 tcaagcccgc cgtgctggag aggcagcgca agagcaaggg ggtgccccgt ggtgaagcct
     5641 aagaacaagc acagcctcag ccacgtccgg cacgacccgg cgcactgtct ggcccccggc
     5701 ctgttccgtg ccctcaagcg gggcgagcgc aagcgcagca agctggacgt gacgtatgac
     5761 tacggtgacg gcaagcggat cgagttcagc ggcccggagc cgctgggcgc tgatgatctg
     5821 cgcatcctgc aagggctggt ggccatggct gggcctaatg gcctagtgct tggcccggaa
     5881 cccaagaccg aaggcggacg gcagctccgg ctgttcctgg aacccaagtg ggaggccgtc
     5941 accgctgatg ccatggtggt caaaggtagc tatcgggcgc tggcaaagga aatcggggca
     6001 gaggtcgata gtggtggggc gctcaagcac atacaggact gcatcgagcg cctttggaag
     6061 gtatccatca tcgcccagaa tggccgcaag cggcaggggt ttcggctgct gtcggagtac
     6121 gccagcgacg aggcggacgg gcgcctgtac gtggccctga accccttgat cgcgcaggcc
     6181 gtcatgggtg gcggccagca tgtgcgcatc agcatggacg aggtgcgggc gctggacagc
     6241 gaaaccgccc gcctgctgca ccagcggctg tgtggctgga tcgaccccgg caaaaccggc
     6301 aaggcttcca tagatgcctt gtgcggctat gtctggccgt cagaggccag tggttcgacc
     6361 atgcgcaagc gccgccagcg ggtgcgcgag gcgttgccgg agctggtcgc gctgggctgg
     6421 acggtaaccg agttcgcggc gggcaagtac gacatcaccc ggcccaaggc ggcaggctga
     6481 ccccccccac tctattgtaa acaagacatt tttatctttt atattcaatg gcttattttc
     6541 ctgctaattg gtaataccat gaaaaatacc atgctcagaa aaggcttaac aatattttga
     6601 aaaattgcct actgagcgct gccgcacagc tccataggcc gttactgaga cgccattcct
     6661 gtagacttct taattaagac gtcagaattc tcgaggcggc cgcatgtgag tctccctata
     6721 gtgagtcgta ttaatttcgc gggcggaacc cctatttgtt tatttttcta aatacattca
     6781 aatatgtatc cgctcatgag tagcaccagg cgtttaaggg caccaataac tgccttaaaa
     6841 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     6901 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     6961 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     7021 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga caaaaaacat
     7081 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     7141 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     7201 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     7261 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
     7321 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     7381 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     7441 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     7501 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     7561 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaaagt ctcattttcg
     7621 ccaaaagttg tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7681 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7741 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7801 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7861 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7921 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7981 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8041 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8101 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8161 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8221 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8281 cggagcctat ggaaaaacgc cagcaatgcg gcctttttac ggttcctggc cttttgctgg
     8341 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8401 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     8461 agcgaggaag c
//
