LOCUS       62056_19535        4556 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb227, complete sequence.
ACCESSION   MW476669
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4556)
  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 4556)
  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 4556)
  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..4556
                     /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"
     CDS             1081..1893
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            2063..2171
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2235..3380)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(3773..4304)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4431..4517)
                     /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 aaaaaaggtc
      841 aaggtctagt cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
      901 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc
      961 caataattac gatttaaatt tgtgtctcaa aatctctgat gttacattgc acaagataaa
     1021 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     1081 atgagccata ttcagcgtga aacgagctgt agccgtccgc gtctgaacag caacatggat
     1141 gcggatctgt atggctataa atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt
     1201 tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc
     1261 gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact ggctgaccga atttatgccg
     1321 ctgccgacca ttaaacattt tattcgcacc ccggatgatg cgtggctgct gaccaccgcg
     1381 attccgggca aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt
     1441 gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg
     1501 tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa caacggcctg
     1561 gtggatgcga gcgattttga tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa
     1621 gaaatgcata aactgctgcc gtttagcccg gatagcgtgg tgacccacgg cgattttagc
     1681 ctggataacc tgattttcga tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg
     1741 ggcattgcgg atcgttatca ggatctggcc attctgtgga actgcctggg cgaatttagc
     1801 ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg ataatccgga tatgaacaaa
     1861 ctgcaatttc atctgatgct ggatgaattt ttctaataat taattggacc gcggtccgcg
     1921 cgttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
     1981 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt
     2041 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
     2101 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
     2161 gaggctggcc gtaggccggc cgcgatgcag gtggctgctg aacccccagc cggaactgac
     2221 cccacaaggc cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc
     2281 aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg
     2341 cgggtggaat ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc
     2401 cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc
     2461 tcccatatga atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc
     2521 aggacctggc aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc
     2581 gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg
     2641 cgcgacaggc attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc
     2701 tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac
     2761 tccaacacct gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag
     2821 gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg
     2881 attttcttgt tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc
     2941 atcgtgtccg gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc
     3001 ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc caggtcctcg
     3061 ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc
     3121 gccaaacctg ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg
     3181 ggcagggcag ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg
     3241 actatcgagc cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg
     3301 atggtttcgg catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc
     3361 cggtcaaacg tccgattcat tcaccctcct tgcgggattg ccccggaatt aattccccgg
     3421 atcgatccgt cgatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     3481 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt
     3541 cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta
     3601 cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     3661 tccggggtca gcaccgtttc tgcggactgg ctttctacgt ggctgccatt tttggggtga
     3721 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc
     3781 gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg
     3841 cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt
     3901 tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac
     3961 agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc
     4021 aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca
     4081 cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg
     4141 ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct
     4201 gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca
     4261 gtgagggcca agttttccgc gaggtatcca caacgccggc ggccctacat ggctctgctg
     4321 tagtgagtgg gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa
     4381 gaagatcctt tgatcttttc tacggcgcgc ccagctgtct agggcggcgg atttgtccta
     4441 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga
     4501 gcctttcgtt ttatttgatg cctttaatta atagctttcg ctaaggatga tttctg
//
