LOCUS       62056_19590        5880 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb657, complete sequence.
ACCESSION   MW476679
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5880)
  AUTHORS   Damalas SG, Batianis C, Martin-Pascual M, de Lorenzo V, Martins Dos 
            Santos VAP.
  TITLE     SEVA 3.1: enabling interoperability of DNA assembly among the SEVA, 
            BioBricks and Type IIS restriction enzyme standards
  JOURNAL   Microb Biotechnol 13 (6), 1793-1806 (2020)
  PUBMED    32710525
REFERENCE   2  (bases 1 to 5880)
  AUTHORS   Damalas SG, Batianis C, Martin Pascual M, de Lorenzo V, Martins dos 
            Santos VAP.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JAN-2021) Laboratory of Systems and Synthetic Biology,
            Wageningen University, Stippeneng 4, Wageningen, Gelderland 6708 WE,
            The Netherlands
REFERENCE   3  (bases 1 to 5880)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb 
            Biotechnol"; date: "2020"; volume: "13"; issue: "6"; pages: 
            "1793-1806"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JAN-2021) Laboratory of Systems and Synthetic Biology, 
            Wageningen University, Stippeneng 4, Wageningen, Gelderland 6708 WE,
            The Netherlands"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      23..57
                     /label=promoter BBa_J23105
                     /note="promoter BBa_J23105"
                     /regulatory_class="promoter"
     regulatory      66..77
                     /label=RBS.20
                     /note="RBS.20"
                     /regulatory_class="ribosome_binding_site"
     CDS             84..797
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    804..824
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      855..949
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        980..1008
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1197..1727
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     oriT            1941..2049
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2063..2911)
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(2901..3737)
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(4251..5219)
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     promoter        complement(5243..5273)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      5329..5723
                     /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)"
     terminator      complement(5755..5841)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gaattcgcgg ccgcttctag agtttacggc tagctcagtc ctaggtacta tgctagctac
       61 tagagtattt aaggctgtac tagatgcgta aaggcgaaga actgttcacg ggcgtagttc
      121 cgattctggt cgagctggac ggcgatgtga acggtcataa gtttagcgtt cgcggtgaag
      181 gtgagggcga cgcgaccaac ggcaaactga ccctgaagtt catctgcacc accggtaaac
      241 tgccggtgcc ttggccgacc ttggtgacga cgttgacgta tggcgtgcag tgttttgcgc
      301 gttatccgga ccacatgaaa caacacgatt tcttcaaatc tgcgatgccg gagggttacg
      361 tccaggagcg taccatttcc ttcaaggatg atggctacta caaaactcgc gcagaggtta
      421 agtttgaagg tgacacgctg gtcaatcgta tcgaattgaa gggtatcgac tttaaagagg
      481 atggtaacat tctgggccat aaactggagt ataacttcaa cagccataat gtttacatta
      541 cggcagacaa gcaaaagaac ggcatcaagg ccaatttcaa gattcgccac aatgttgagg
      601 acggtagcgt ccaactggcc gaccattacc agcagaacac cccaattggt gacggtccgg
      661 ttttgctgcc ggataatcac tatctgagca cccaaagcgt gctgagcaaa gatccgaacg
      721 aaaaacgtga tcacatggtc ctgctggaat ttgtgaccgc tgcgggcatc acccacggta
      781 tggatgaact gtacaaataa taatactagt agcggccgct gcagtccggc aaaaaagggc
      841 aaggtctagt cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
      901 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc
      961 caataattac gatttaaatt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     1021 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     1081 agtggcggtt ttcatggctt gttatgactg tttttttgta cagcctatgc ctcgggcatc
     1141 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     1201 tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct
     1261 caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg
     1321 ctgctcttga tcttttcggt cgtgagttcg gagacgtagc cacctactcc caacatcagc
     1381 cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct
     1441 tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccaag tttgagcagc
     1501 cgcgtagtga gatctatatc tatgatctcg cagtctccgg agagcaccgg aggcagggca
     1561 ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga
     1621 tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca
     1681 tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc
     1741 aagccgagat cggcttcccg gccgcggagt tgttcggtaa attggacaac ggtccgcgcg
     1801 ttgtcctttt ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat
     1861 ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg
     1921 tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt
     1981 tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga
     2041 ggctggccgt aggccggcct cagcctgccg ccttgggccg ggtgatgtcg tacttgcccg
     2101 ccgcgaactc ggttaccgtc cagcccagcg cgaccagctc cggcaacgcc tcgcgcaccc
     2161 gctggcggcg cttgcgcatg gtcgaaccac tggcctctga cggccagaca tagccgcaca
     2221 aggtatctat ggaagccttg ccggttttgc cggggtcgat ccagccacac agccgctggt
     2281 gcagcaggcg ggcggtttcg ctgtccagcg cccgcacctc gtccatgctg atgcgcacat
     2341 gctggccgcc acccatgacg gcctgcgcga tcaaggggtt cagggccacg tacaggcgcc
     2401 cgtccgcctc gtcgctggcg tactccgaca gcagccgaaa cccctgccgc ttgcggccat
     2461 tctgggcgat gatggatacc ttccaaaggc gctcgatgca gtcctgtatg tgcttgagcg
     2521 ccccaccact atcgacctct gccccgattt cctttgccag cgcccgatag ctacctttga
     2581 ccacatggca ttcagcggtg acggcctccc acttgggttc caggaacagc cggagctgcc
     2641 gtccgccttc ggtcttgggt tccgggccaa gcactaggcc attaggccca gccatggcca
     2701 ccagcccttg caggatgcgc agatcatcag cgcccagcgg ctccgggccg ctgaactcga
     2761 tccgcttgcc gtcgccgtag tcatacgtca cgtccagctt gctgcgcttg cgctcgcccc
     2821 gcttgagggc acggaacagg ccgggggcca gacagtgcgc cgggtcgtgc cggacgtggc
     2881 tgaggctgtg cttgttctta ggcttcacca cggggcaccc ccttgctctt gcgctgcctc
     2941 tccagcacgg cgggcttgag caccccgccg tcatgccgcc tgaaccaccg atcagcgaac
     3001 ggtgcgccat agttggcctt gctcacaccg aagcggacga agaaccggcg ctggtcgtcg
     3061 tccacacccc attcctcggc ctcggcgctg gtcatgctcg acaggtagga ctgccagcgg
     3121 atgttatcga ccagtaccga gctgccccgg ctggcctgct gctggtcgcc tgcgcccatc
     3181 atggccgcgc ccttgctggc atggtgcagg aacacgatag agcacccggt atcggcggcg
     3241 atggcctcca tgcgaccgat gacctgggcc atggggccgc tggcgttttc ttcctcgatg
     3301 tggaaccggc gcagcgtgtc cagcaccatc aggcggcggc cctcggcggc gcgcttgagg
     3361 ccgtcgaacc actccggggc catgatgttg ggcaggctgc cgatcagcgg ctggatcagc
     3421 aggccgtcag ccacggcttg ccgttcctcg gcgctgaggt gcgccccaag ggcgtgcagg
     3481 cggtgatgaa tggcggtggg cgggtcttcg gcgggcaggt agatcaccgg gccggtgggc
     3541 agttcgccca cctccagcag atccggcccg cctgcaatct gtgcggccag ttgcagggcc
     3601 agcatggatt taccggcacc accgggcgac accagcgccc cgaccgtacc ggccaccatg
     3661 ttgggcaaaa cgtagtccag cggtggcggc gctgctgcga acgcctccag aatattgata
     3721 ggcttatggg tagccattga ttgcctcctt tgcaggcagt tggtggttag gcgctggcgg
     3781 ggtcactacc cccgccctgc gccgctctga gttcttccag gcactcgcgc agcgcctcgt
     3841 attcgtcgtc ggtcagccag aacttgcgct gacgcatccc tttggccttc atgcgctcgg
     3901 catatcgcgc ttggcgtaca gcgtcagggc tggccagcag gtcgccggtc tgcttgtcct
     3961 tttggtcttt catatcagtc accgagaaac ttgccggggc cgaaaggctt gtcttcgcgg
     4021 aacaaggaca aggtgcagcc gtcaaggtta aggctggcca tatcagcgac tgaaaagcgg
     4081 ccagcctcgg ccttgtttga cgtataacca aagccaccgg gcaaccaata gcccttgtca
     4141 cttttgatca ggtagaccga ccctgaagcg cttttttcgt attccataaa acccccttct
     4201 gtgcgtgagt actcatagta taacaggcgt gagtaccaac gcaagcacta catgctgaaa
     4261 tctggcccgc ccctgtccat gcctcgctgg cggggtgccg gtgcccgtgc cagctcggcc
     4321 cgcgcaagct ggacgctggg cagacccatg accttgctga cggtgcgctc gatgtaatcc
     4381 gcttcgtggc cgggcttgcg ctctgccagc gctgggctgg cctcggccat ggccttgccg
     4441 atttcctcgg cactgcggcc ccggctggcc agcttctgcg cggcgataaa gtcgcacttg
     4501 ctgaggtcat caccgaagcg cttgaccagc ccggccatct cgctgcggta ctcgtccagc
     4561 gccgtgcgcc ggtggcggct aagctgccgc tcgggcagtt cgaggctggc cagcctgcgg
     4621 gccttctcct gctgccgctg ggcctgctcg atctgctggc cagcctgctg caccagcgcc
     4681 gggccagcgg tggcggtctt gcccttggat tcacgcagca gcacccacgg ctgataaccg
     4741 gcgcgggtgg tgtgcttgtc cttgcggttg gtgaagcccg ccaagcggcc atagtggcgg
     4801 ctgtcggcgc tggccgggtc ggcgtcgtac tcgctggcca gcgtccgggc aatctgcccc
     4861 cgaagttcac cgcctgcggc gtcggccacc ttgacccatg cctgatagtt cttcgggctg
     4921 gtttccacta ccagggcagg ctcccggccc tcggctttca tgtcatccag gtcaaactcg
     4981 ctgaggtcgt ccaccagcac cagaccatgc cgctcctgct cggcgggcct gatatacacg
     5041 tcattgccct gggcattcat ccgcttgagc catggcgtgt tctggagcac ttcggcggct
     5101 gaccattccc ggttcatcat ctggccggtg gtggcgtccc tgacgccgat atcgaagcgc
     5161 tcacagccca tggccttgag ctgtcggcct atggcctgca aagtcctgtc gttcttcatc
     5221 gggccaccaa gcgattccca cacattatac gagccggaag cataaagtgt aaagcctaga
     5281 tccgaaggat gagccgggct gaatgatcga ccgagacagg ccctgcgggg ctgcacacgc
     5341 gcccccaccc ttcgggtagg gggaaaggcc gctaaagcgg ctaaaagcgc tccagcgtat
     5401 ttctgcgggg tttggtgtgg ggtttagcgg gctttgcccg cctttccccc tgccgcgcag
     5461 cggtggggcg gtgtgtagcc tagcgcagcg aatagaccag ctatccggcc tctggccggg
     5521 catattgggc aagggcagca gcgccccaca agggcgctga taaccgcgcc tagtggatta
     5581 ttcttagata atcatggatg gatttttcca acaccccgcc agcccccgcc cctgctgggt
     5641 ttgcaggttt gggggcgtga cagttattgc aggggttcgt gacagttatt gcaggggggc
     5701 gtgacagtta ttgcaggggt tcgtgacagt tagggcgcgc ctctagggcg gcggatttgt
     5761 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     5821 ctgagccttt cgttttattt gatgccttta attaatagct ttcgctaagg atgatttctg
//
