LOCUS       62056_19555        4616 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb427, complete sequence.
ACCESSION   MW476673
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4616)
  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 4616)
  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 4616)
  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..4616
                     /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             1185..1973
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     oriT            2122..2230
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2294..3439)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     rep_origin      complement(3832..4363)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4490..4576)
                     /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 gatttacgta tttaaatgaa ccttgaccga acgcagcggt ggtaacggcg
     1021 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     1081 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     1141 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga
     1201 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     1261 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     1321 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     1381 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     1441 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     1501 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     1561 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     1621 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     1681 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     1741 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     1801 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     1861 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1921 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataagaca
     1981 attgtccttt tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata
     2041 tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg
     2101 gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg
     2161 ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg
     2221 aggctggccg taggccggcc gcgatgcagg tggctgctga acccccagcc ggaactgacc
     2281 ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca
     2341 ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc
     2401 gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc
     2461 ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct
     2521 cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca
     2581 ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg
     2641 acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc
     2701 gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct
     2761 ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact
     2821 ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg
     2881 tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga
     2941 ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca
     3001 tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct
     3061 tcgtgtgttt cagcaacgcg gcctgcttgg cttcgctgac ctgttttgcc aggtcctcgc
     3121 cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg
     3181 ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc gatggcgcgg
     3241 gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga
     3301 ctatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga
     3361 tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc
     3421 ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccggaatta attccccgga
     3481 tcgatccgtc gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     3541 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
     3601 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     3661 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     3721 ccggggtcag caccgtttct gcggactggc tttctacgtg gctgccattt ttggggtgag
     3781 gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt tagcgggccg
     3841 ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc
     3901 agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt
     3961 agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca
     4021 gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca
     4081 aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac
     4141 ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc
     4201 cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg
     4261 tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag
     4321 tgagggccaa gttttccgcg aggtatccac aacgccggcg gccctacatg gctctgctgt
     4381 agtgagtggg ttgcgctccg gcagcggtcc tgatcccccg cagaaaaaaa ggatctcaag
     4441 aagatccttt gatcttttct acggcgcgcc cagctgtcta gggcggcgga tttgtcctac
     4501 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     4561 cctttcgttt tatttgatgc cttttaatta atagctttcg ctaaggatga tttctg
//
