LOCUS       62056_19550        6008 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb257, complete sequence.
ACCESSION   MW476672
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6008)
  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 6008)
  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 6008)
  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..6008
                     /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(2185..3033)
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(3023..3859)
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(4373..5341)
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     promoter        complement(5365..5395)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      5451..5845
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(5883..5969)
                     /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 ctcagcctgc cgccttgggc cgggtgatgt cgtacttgcc
     2221 cgccgcgaac tcggttaccg tccagcccag cgcgaccagc tccggcaacg cctcgcgcac
     2281 ccgctggcgg cgcttgcgca tggtcgaacc actggcctct gacggccaga catagccgca
     2341 caaggtatct atggaagcct tgccggtttt gccggggtcg atccagccac acagccgctg
     2401 gtgcagcagg cgggcggttt cgctgtccag cgcccgcacc tcgtccatgc tgatgcgcac
     2461 atgctggccg ccacccatga cggcctgcgc gatcaagggg ttcagggcca cgtacaggcg
     2521 cccgtccgcc tcgtcgctgg cgtactccga cagcagccga aacccctgcc gcttgcggcc
     2581 attctgggcg atgatggata ccttccaaag gcgctcgatg cagtcctgta tgtgcttgag
     2641 cgccccacca ctatcgacct ctgccccgat ttcctttgcc agcgcccgat agctaccttt
     2701 gaccacatgg cattcagcgg tgacggcctc ccacttgggt tccaggaaca gccggagctg
     2761 ccgtccgcct tcggtcttgg gttccgggcc aagcactagg ccattaggcc cagccatggc
     2821 caccagccct tgcaggatgc gcagatcatc agcgcccagc ggctccgggc cgctgaactc
     2881 gatccgcttg ccgtcgccgt agtcatacgt cacgtccagc ttgctgcgct tgcgctcgcc
     2941 ccgcttgagg gcacggaaca ggccgggggc cagacagtgc gccgggtcgt gccggacgtg
     3001 gctgaggctg tgcttgttct taggcttcac cacggggcac ccccttgctc ttgcgctgcc
     3061 tctccagcac ggcgggcttg agcaccccgc cgtcatgccg cctgaaccac cgatcagcga
     3121 acggtgcgcc atagttggcc ttgctcacac cgaagcggac gaagaaccgg cgctggtcgt
     3181 cgtccacacc ccattcctcg gcctcggcgc tggtcatgct cgacaggtag gactgccagc
     3241 ggatgttatc gaccagtacc gagctgcccc ggctggcctg ctgctggtcg cctgcgccca
     3301 tcatggccgc gcccttgctg gcatggtgca ggaacacgat agagcacccg gtatcggcgg
     3361 cgatggcctc catgcgaccg atgacctggg ccatggggcc gctggcgttt tcttcctcga
     3421 tgtggaaccg gcgcagcgtg tccagcacca tcaggcggcg gccctcggcg gcgcgcttga
     3481 ggccgtcgaa ccactccggg gccatgatgt tgggcaggct gccgatcagc ggctggatca
     3541 gcaggccgtc agccacggct tgccgttcct cggcgctgag gtgcgcccca agggcgtgca
     3601 ggcggtgatg aatggcggtg ggcgggtctt cggcgggcag gtagatcacc gggccggtgg
     3661 gcagttcgcc cacctccagc agatccggcc cgcctgcaat ctgtgcggcc agttgcaggg
     3721 ccagcatgga tttaccggca ccaccgggcg acaccagcgc cccgaccgta ccggccacca
     3781 tgttgggcaa aacgtagtcc agcggtggcg gcgctgctgc gaacgcctcc agaatattga
     3841 taggcttatg ggtagccatt gattgcctcc tttgcaggca gttggtggtt aggcgctggc
     3901 ggggtcacta cccccgccct gcgccgctct gagttcttcc aggcactcgc gcagcgcctc
     3961 gtattcgtcg tcggtcagcc agaacttgcg ctgacgcatc cctttggcct tcatgcgctc
     4021 ggcatatcgc gcttggcgta cagcgtcagg gctggccagc aggtcgccgg tctgcttgtc
     4081 cttttggtct ttcatatcag tcaccgagaa acttgccggg gccgaaaggc ttgtcttcgc
     4141 ggaacaagga caaggtgcag ccgtcaaggt taaggctggc catatcagcg actgaaaagc
     4201 ggccagcctc ggccttgttt gacgtataac caaagccacc gggcaaccaa tagcccttgt
     4261 cacttttgat caggtagacc gaccctgaag cgcttttttc gtattccata aaaccccctt
     4321 ctgtgcgtga gtactcatag tataacaggc gtgagtacca acgcaagcac tacatgctga
     4381 aatctggccc gcccctgtcc atgcctcgct ggcggggtgc cggtgcccgt gccagctcgg
     4441 cccgcgcaag ctggacgctg ggcagaccca tgaccttgct gacggtgcgc tcgatgtaat
     4501 ccgcttcgtg gccgggcttg cgctctgcca gcgctgggct ggcctcggcc atggccttgc
     4561 cgatttcctc ggcactgcgg ccccggctgg ccagcttctg cgcggcgata aagtcgcact
     4621 tgctgaggtc atcaccgaag cgcttgacca gcccggccat ctcgctgcgg tactcgtcca
     4681 gcgccgtgcg ccggtggcgg ctaagctgcc gctcgggcag ttcgaggctg gccagcctgc
     4741 gggccttctc ctgctgccgc tgggcctgct cgatctgctg gccagcctgc tgcaccagcg
     4801 ccgggccagc ggtggcggtc ttgcccttgg attcacgcag cagcacccac ggctgataac
     4861 cggcgcgggt ggtgtgcttg tccttgcggt tggtgaagcc cgccaagcgg ccatagtggc
     4921 ggctgtcggc gctggccggg tcggcgtcgt actcgctggc cagcgtccgg gcaatctgcc
     4981 cccgaagttc accgcctgcg gcgtcggcca ccttgaccca tgcctgatag ttcttcgggc
     5041 tggtttccac taccagggca ggctcccggc cctcggcttt catgtcatcc aggtcaaact
     5101 cgctgaggtc gtccaccagc accagaccat gccgctcctg ctcggcgggc ctgatataca
     5161 cgtcattgcc ctgggcattc atccgcttga gccatggcgt gttctggagc acttcggcgg
     5221 ctgaccattc ccggttcatc atctggccgg tggtggcgtc cctgacgccg atatcgaagc
     5281 gctcacagcc catggccttg agctgtcggc ctatggcctg caaagtcctg tcgttcttca
     5341 tcgggccacc aagcgattcc cacacattat acgagccgga agcataaagt gtaaagccta
     5401 gatccgaagg atgagccggg ctgaatgatc gaccgagaca ggccctgcgg ggctgcacac
     5461 gcgcccccac ccttcgggta gggggaaagg ccgctaaagc ggctaaaagc gctccagcgt
     5521 atttctgcgg ggtttggtgt ggggtttagc gggctttgcc cgcctttccc cctgccgcgc
     5581 agcggtgggg cggtgtgtag cctagcgcag cgaatagacc agctatccgg cctctggccg
     5641 ggcatattgg gcaagggcag cagcgcccca caagggcgct gataaccgcg cctagtggat
     5701 tattcttaga taatcatgga tggatttttc caacaccccg ccagcccccg cccctgctgg
     5761 gtttgcaggt ttgggggcgt gacagttatt gcaggggttc gtgacagtta ttgcaggggg
     5821 gcgtgacagt tattgcaggg gttcgtgaca gttagggcgc gcccagctgt ctagggcggc
     5881 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     5941 tctttcgact gagcctttcg ttttatttga tgcctttaat taatagcttt cgctaaggat
     6001 gatttctg
//
