LOCUS       62056_19585        4181 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb647, complete sequence.
ACCESSION   MW476678
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4181)
  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 4181)
  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 4181)
  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..4181
                     /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"
     rep_origin      2143..2731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2788..3618)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      3632..3983
                     /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(4056..4142)
                     /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 tgaaataagc 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 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2101 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2161 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2221 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2281 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2341 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2401 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2461 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2521 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggca
     2581 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2641 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2701 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggccg tgaaaggcag
     2761 gccggtccgt ggtggccacg gcctctaggc cagatccagc ggcatctggg ttagtcgagc
     2821 gcgggccgct tcccatgtct caccagggcg agcctgtttc gcgatctcag catctgaaat
     2881 cttcccggcc ttgcgcttcg ctggggcctt acccaccgcc ttggcgggct tcttcggtcc
     2941 aaaactgaac aacagatgtg tgaccttgcg cccggtcttt cgctgcgccc actccacctg
     3001 tagcgggctg tgctcgttga tctgcgtcac ggctggatca agcactcgca acttgaagtc
     3061 cttgatcgag ggataccggc cttccagttg aaaccacttt cgcagctggt caatttctat
     3121 ttcgcgctgg ccgatgctgt cccattgcat gagcagctcg taaagcctga tcgcgtgggt
     3181 gctgtccatc ttggccacgt cagccaaggc gtatttggtg aactgtttgg tgagttccgt
     3241 caggtacggc agcatgtctt tggtgaacct gagttctaca cggccctcac cctcccggta
     3301 gatgattgtt tgcacccagc cggtaatcat cacactcggt cttttcccct tgccattggg
     3361 ctcttgggtt aaccggactt cccgccgttt caggcgcagg gccgcttctt tgagctggtt
     3421 gtaggaagat tcgataggga cacccgccat cgtcgctatg tcctccgccg tcactgaata
     3481 catcacttca tcggtgacag gctcgctcct cttcacctgg ctaatacagg ccagaacgat
     3541 ccgctgttcc tgaacactga ggcgatacgc ggcctcgacc agggcattgc ttttgtaaac
     3601 cattgggggt gaggccacgt tcgacattcc ttgtgtataa ggggacactg tatctgcgtc
     3661 ccacaataca acaaatccgt ccctttacaa caacaaatcc gtcccttctt aacaacaaat
     3721 ccgtccctta atggcaacaa atccgtccct ttttaaactc tacaggccac ggattacgtg
     3781 gcctgtagac gtcctaaaag gtttaaaagg gaaaaggaag aaaagggtgg aaacgcaaaa
     3841 aacgcaccac tacgtggccc cgttggggcc gcatttgtgc ccctgaaggg gcgggggagg
     3901 cgtctgggca atccccgttt taccagtccc ctatcgccgc ctgagagggc gcaggaagcg
     3961 agtaatcagg gtatcgaggc ggattcaccc ttggcgtcca accagcggca ccagcggcgc
     4021 ctgagagggg cgcgcccagc tgtctagggc ggcggatttg tcctactcag gagagcgttc
     4081 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     4141 tgatgccttt aattaatagc tttcgctaag gatgatttct g
//
