LOCUS       62056_19545        4303 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb247, complete sequence.
ACCESSION   MW476671
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4303)
  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 4303)
  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 4303)
  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..4303
                     /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"
     rep_origin      2265..2853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2910..3740)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      3754..4105
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      complement(4178..4264)
                     /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 cgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2701 catccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc cgtgaaaggc
     2881 aggccggtcc gtggtggcca cggcctctag gccagatcca gcggcatctg ggttagtcga
     2941 gcgcgggccg cttcccatgt ctcaccaggg cgagcctgtt tcgcgatctc agcatctgaa
     3001 atcttcccgg ccttgcgctt cgctggggcc ttacccaccg ccttggcggg cttcttcggt
     3061 ccaaaactga acaacagatg tgtgaccttg cgcccggtct ttcgctgcgc ccactccacc
     3121 tgtagcgggc tgtgctcgtt gatctgcgtc acggctggat caagcactcg caacttgaag
     3181 tccttgatcg agggataccg gccttccagt tgaaaccact ttcgcagctg gtcaatttct
     3241 atttcgcgct ggccgatgct gtcccattgc atgagcagct cgtaaagcct gatcgcgtgg
     3301 gtgctgtcca tcttggccac gtcagccaag gcgtatttgg tgaactgttt ggtgagttcc
     3361 gtcaggtacg gcagcatgtc tttggtgaac ctgagttcta cacggccctc accctcccgg
     3421 tagatgattg tttgcaccca gccggtaatc atcacactcg gtcttttccc cttgccattg
     3481 ggctcttggg ttaaccggac ttcccgccgt ttcaggcgca gggccgcttc tttgagctgg
     3541 ttgtaggaag attcgatagg gacacccgcc atcgtcgcta tgtcctccgc cgtcactgaa
     3601 tacatcactt catcggtgac aggctcgctc ctcttcacct ggctaataca ggccagaacg
     3661 atccgctgtt cctgaacact gaggcgatac gcggcctcga ccagggcatt gcttttgtaa
     3721 accattgggg gtgaggccac gttcgacatt ccttgtgtat aaggggacac tgtatctgcg
     3781 tcccacaata caacaaatcc gtccctttac aacaacaaat ccgtcccttc ttaacaacaa
     3841 atccgtccct taatggcaac aaatccgtcc ctttttaaac tctacaggcc acggattacg
     3901 tggcctgtag acgtcctaaa aggtttaaaa gggaaaagga agaaaagggt ggaaacgcaa
     3961 aaaacgcacc actacgtggc cccgttgggg ccgcatttgt gcccctgaag gggcggggga
     4021 ggcgtctggg caatccccgt tttaccagtc ccctatcgcc gcctgagagg gcgcaggaag
     4081 cgagtaatca gggtatcgag gcggattcac ccttggcgtc caaccagcgg caccagcggc
     4141 gcctgagagg ggcgcgccca gctgtctagg gcggcggatt tgtcctactc aggagagcgt
     4201 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     4261 tttgatgcct ttaattaata gctttcgcta aggatgattt ctg
//
