LOCUS       62056_19575        4434 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb627, complete sequence.
ACCESSION   MW476676
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4434)
  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 4434)
  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 4434)
  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..4434
                     /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(2113..3258)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(3651..4182)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4309..4395)
                     /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 aggccggccg cgatgcaggt ggctgctgaa cccccagccg gaactgaccc
     2101 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     2161 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     2221 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     2281 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     2341 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     2401 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     2461 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     2521 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     2581 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     2641 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     2701 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     2761 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     2821 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     2881 cgtgtgtttc agcaacgcgg cctgcttggc ttcgctgacc tgttttgcca ggtcctcgcc
     2941 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     3001 caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg
     3061 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     3121 tatcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     3181 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     3241 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccggaattaa ttccccggat
     3301 cgatccgtcg atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     3361 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     3421 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     3481 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     3541 cggggtcagc accgtttctg cggactggct ttctacgtgg ctgccatttt tggggtgagg
     3601 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg
     3661 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca
     3721 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta
     3781 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag
     3841 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa
     3901 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact
     3961 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc
     4021 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt
     4081 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt
     4141 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccctacatgg ctctgctgta
     4201 gtgagtgggt tgcgctccgg cagcggtcct gatcccccgc agaaaaaaag gatctcaaga
     4261 agatcctttg atcttttcta cggcgcgccc agctgtctag ggcggcggat ttgtcctact
     4321 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
     4381 ctttcgtttt atttgatgcc tttaattaat agctttcgct aaggatgatt tctg
//
