LOCUS       62056_19530        6103 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb157, complete sequence.
ACCESSION   MW476668
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6103)
  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 6103)
  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 6103)
  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..6103
                     /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        1022..1113
                     /label=AmpR promoter
     CDS             1114..1971
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      1978..2017
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            2164..2272
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2286..3134)
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(3124..3960)
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(4474..5442)
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     promoter        complement(5466..5496)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      5552..5946
                     /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(5978..6064)
                     /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 agtagcccgc ctaatgagcg ggcttttttt taattcccct
     1021 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1081 taaatgcttc aataatattg aaaaaggaag agtatgagca ttcagcattt tcgtgtggcg
     1141 ctgattccgt tttttgcggc gttttgcctg ccggtgtttg cgcatccgga aaccctggtg
     1201 aaagtgaaag atgcggaaga tcaactgggt gcgcgcgtgg gctatattga actggatctg
     1261 aacagcggca aaattctgga atcttttcgt ccggaagaac gttttccgat gatgagcacc
     1321 tttaaagtgc tgctgtgcgg tgcggttctg agccgtgtgg atgcgggcca ggaacaactg
     1381 ggccgtcgta ttcattatag ccagaacgat ctggtggaat atagcccggt gaccgaaaaa
     1441 catctgaccg atggcatgac cgtgcgtgaa ctgtgcagcg cggcgattac catgagcgat
     1501 aacaccgcgg cgaacctgct gctgacgacc attggcggtc cgaaagaact gaccgcgttt
     1561 ctgcataaca tgggcgatca tgtgacccgt ctggatcgtt gggaaccgga actgaacgaa
     1621 gcgattccga acgatgaacg tgataccacc atgccggcag caatggcgac caccctgcgt
     1681 aaactgctga cgggtgagct gctgaccctg gcaagccgcc agcaactgat tgattggatg
     1741 gaagcggata aagtggcggg tccgctgctg cgtagcgcgc tgccggctgg ctggtttatt
     1801 gcggataaaa gcggtgcggg cgaacgtggc agccgtggca ttattgcggc gctgggcccg
     1861 gatggtaaac cgagccgtat tgtggtgatt tataccaccg gcagccaggc gacgatggat
     1921 gaacgtaacc gtcagattgc ggaaattggc gcgagcctga ttaaacattg gtaaaccgat
     1981 acaattaaag gctccttttg gagccttttt ttttggacga ccccggtcct tttccgctgc
     2041 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     2101 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     2161 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     2221 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg
     2281 cctcagcctg ccgccttggg ccgggtgatg tcgtacttgc ccgccgcgaa ctcggttacc
     2341 gtccagccca gcgcgaccag ctccggcaac gcctcgcgca cccgctggcg gcgcttgcgc
     2401 atggtcgaac cactggcctc tgacggccag acatagccgc acaaggtatc tatggaagcc
     2461 ttgccggttt tgccggggtc gatccagcca cacagccgct ggtgcagcag gcgggcggtt
     2521 tcgctgtcca gcgcccgcac ctcgtccatg ctgatgcgca catgctggcc gccacccatg
     2581 acggcctgcg cgatcaaggg gttcagggcc acgtacaggc gcccgtccgc ctcgtcgctg
     2641 gcgtactccg acagcagccg aaacccctgc cgcttgcggc cattctgggc gatgatggat
     2701 accttccaaa ggcgctcgat gcagtcctgt atgtgcttga gcgccccacc actatcgacc
     2761 tctgccccga tttcctttgc cagcgcccga tagctacctt tgaccacatg gcattcagcg
     2821 gtgacggcct cccacttggg ttccaggaac agccggagct gccgtccgcc ttcggtcttg
     2881 ggttccgggc caagcactag gccattaggc ccagccatgg ccaccagccc ttgcaggatg
     2941 cgcagatcat cagcgcccag cggctccggg ccgctgaact cgatccgctt gccgtcgccg
     3001 tagtcatacg tcacgtccag cttgctgcgc ttgcgctcgc cccgcttgag ggcacggaac
     3061 aggccggggg ccagacagtg cgccgggtcg tgccggacgt ggctgaggct gtgcttgttc
     3121 ttaggcttca ccacggggca cccccttgct cttgcgctgc ctctccagca cggcgggctt
     3181 gagcaccccg ccgtcatgcc gcctgaacca ccgatcagcg aacggtgcgc catagttggc
     3241 cttgctcaca ccgaagcgga cgaagaaccg gcgctggtcg tcgtccacac cccattcctc
     3301 ggcctcggcg ctggtcatgc tcgacaggta ggactgccag cggatgttat cgaccagtac
     3361 cgagctgccc cggctggcct gctgctggtc gcctgcgccc atcatggccg cgcccttgct
     3421 ggcatggtgc aggaacacga tagagcaccc ggtatcggcg gcgatggcct ccatgcgacc
     3481 gatgacctgg gccatggggc cgctggcgtt ttcttcctcg atgtggaacc ggcgcagcgt
     3541 gtccagcacc atcaggcggc ggccctcggc ggcgcgcttg aggccgtcga accactccgg
     3601 ggccatgatg ttgggcaggc tgccgatcag cggctggatc agcaggccgt cagccacggc
     3661 ttgccgttcc tcggcgctga ggtgcgcccc aagggcgtgc aggcggtgat gaatggcggt
     3721 gggcgggtct tcggcgggca ggtagatcac cgggccggtg ggcagttcgc ccacctccag
     3781 cagatccggc ccgcctgcaa tctgtgcggc cagttgcagg gccagcatgg atttaccggc
     3841 accaccgggc gacaccagcg ccccgaccgt accggccacc atgttgggca aaacgtagtc
     3901 cagcggtggc ggcgctgctg cgaacgcctc cagaatattg ataggcttat gggtagccat
     3961 tgattgcctc ctttgcaggc agttggtggt taggcgctgg cggggtcact acccccgccc
     4021 tgcgccgctc tgagttcttc caggcactcg cgcagcgcct cgtattcgtc gtcggtcagc
     4081 cagaacttgc gctgacgcat ccctttggcc ttcatgcgct cggcatatcg cgcttggcgt
     4141 acagcgtcag ggctggccag caggtcgccg gtctgcttgt ccttttggtc tttcatatca
     4201 gtcaccgaga aacttgccgg ggccgaaagg cttgtcttcg cggaacaagg acaaggtgca
     4261 gccgtcaagg ttaaggctgg ccatatcagc gactgaaaag cggccagcct cggccttgtt
     4321 tgacgtataa ccaaagccac cgggcaacca atagcccttg tcacttttga tcaggtagac
     4381 cgaccctgaa gcgctttttt cgtattccat aaaaccccct tctgtgcgtg agtactcata
     4441 gtataacagg cgtgagtacc aacgcaagca ctacatgctg aaatctggcc cgcccctgtc
     4501 catgcctcgc tggcggggtg ccggtgcccg tgccagctcg gcccgcgcaa gctggacgct
     4561 gggcagaccc atgaccttgc tgacggtgcg ctcgatgtaa tccgcttcgt ggccgggctt
     4621 gcgctctgcc agcgctgggc tggcctcggc catggccttg ccgatttcct cggcactgcg
     4681 gccccggctg gccagcttct gcgcggcgat aaagtcgcac ttgctgaggt catcaccgaa
     4741 gcgcttgacc agcccggcca tctcgctgcg gtactcgtcc agcgccgtgc gccggtggcg
     4801 gctaagctgc cgctcgggca gttcgaggct ggccagcctg cgggccttct cctgctgccg
     4861 ctgggcctgc tcgatctgct ggccagcctg ctgcaccagc gccgggccag cggtggcggt
     4921 cttgcccttg gattcacgca gcagcaccca cggctgataa ccggcgcggg tggtgtgctt
     4981 gtccttgcgg ttggtgaagc ccgccaagcg gccatagtgg cggctgtcgg cgctggccgg
     5041 gtcggcgtcg tactcgctgg ccagcgtccg ggcaatctgc ccccgaagtt caccgcctgc
     5101 ggcgtcggcc accttgaccc atgcctgata gttcttcggg ctggtttcca ctaccagggc
     5161 aggctcccgg ccctcggctt tcatgtcatc caggtcaaac tcgctgaggt cgtccaccag
     5221 caccagacca tgccgctcct gctcggcggg cctgatatac acgtcattgc cctgggcatt
     5281 catccgcttg agccatggcg tgttctggag cacttcggcg gctgaccatt cccggttcat
     5341 catctggccg gtggtggcgt ccctgacgcc gatatcgaag cgctcacagc ccatggcctt
     5401 gagctgtcgg cctatggcct gcaaagtcct gtcgttcttc atcgggccac caagcgattc
     5461 ccacacatta tacgagccgg aagcataaag tgtaaagcct agatccgaag gatgagccgg
     5521 gctgaatgat cgaccgagac aggccctgcg gggctgcaca cgcgccccca cccttcgggt
     5581 agggggaaag gccgctaaag cggctaaaag cgctccagcg tatttctgcg gggtttggtg
     5641 tggggtttag cgggctttgc ccgcctttcc ccctgccgcg cagcggtggg gcggtgtgta
     5701 gcctagcgca gcgaatagac cagctatccg gcctctggcc gggcatattg ggcaagggca
     5761 gcagcgcccc acaagggcgc tgataaccgc gcctagtgga ttattcttag ataatcatgg
     5821 atggattttt ccaacacccc gccagccccc gcccctgctg ggtttgcagg tttgggggcg
     5881 tgacagttat tgcaggggtt cgtgacagtt attgcagggg ggcgtgacag ttattgcagg
     5941 ggttcgtgac agttagggcg cgcctctagg gcggcggatt tgtcctactc aggagagcgt
     6001 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     6061 tttgatgcct ttaattaata gctttcgcta aggatgattt ctg
//
