LOCUS       62056_19570        6067 bp DNA     circular SYN 27-FEB-2021
DEFINITION  Cloning vector pSEVAb457, complete sequence.
ACCESSION   MW476675
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6067)
  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 6067)
  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 6067)
  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..6067
                     /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(2244..3092)
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(3082..3918)
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             complement(4432..5400)
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     promoter        complement(5424..5454)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      5510..5904
                     /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(5942..6028)
                     /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 tcagcctgcc gccttgggcc gggtgatgtc gtacttgccc
     2281 gccgcgaact cggttaccgt ccagcccagc gcgaccagct ccggcaacgc ctcgcgcacc
     2341 cgctggcggc gcttgcgcat ggtcgaacca ctggcctctg acggccagac atagccgcac
     2401 aaggtatcta tggaagcctt gccggttttg ccggggtcga tccagccaca cagccgctgg
     2461 tgcagcaggc gggcggtttc gctgtccagc gcccgcacct cgtccatgct gatgcgcaca
     2521 tgctggccgc cacccatgac ggcctgcgcg atcaaggggt tcagggccac gtacaggcgc
     2581 ccgtccgcct cgtcgctggc gtactccgac agcagccgaa acccctgccg cttgcggcca
     2641 ttctgggcga tgatggatac cttccaaagg cgctcgatgc agtcctgtat gtgcttgagc
     2701 gccccaccac tatcgacctc tgccccgatt tcctttgcca gcgcccgata gctacctttg
     2761 accacatggc attcagcggt gacggcctcc cacttgggtt ccaggaacag ccggagctgc
     2821 cgtccgcctt cggtcttggg ttccgggcca agcactaggc cattaggccc agccatggcc
     2881 accagccctt gcaggatgcg cagatcatca gcgcccagcg gctccgggcc gctgaactcg
     2941 atccgcttgc cgtcgccgta gtcatacgtc acgtccagct tgctgcgctt gcgctcgccc
     3001 cgcttgaggg cacggaacag gccgggggcc agacagtgcg ccgggtcgtg ccggacgtgg
     3061 ctgaggctgt gcttgttctt aggcttcacc acggggcacc cccttgctct tgcgctgcct
     3121 ctccagcacg gcgggcttga gcaccccgcc gtcatgccgc ctgaaccacc gatcagcgaa
     3181 cggtgcgcca tagttggcct tgctcacacc gaagcggacg aagaaccggc gctggtcgtc
     3241 gtccacaccc cattcctcgg cctcggcgct ggtcatgctc gacaggtagg actgccagcg
     3301 gatgttatcg accagtaccg agctgccccg gctggcctgc tgctggtcgc ctgcgcccat
     3361 catggccgcg cccttgctgg catggtgcag gaacacgata gagcacccgg tatcggcggc
     3421 gatggcctcc atgcgaccga tgacctgggc catggggccg ctggcgtttt cttcctcgat
     3481 gtggaaccgg cgcagcgtgt ccagcaccat caggcggcgg ccctcggcgg cgcgcttgag
     3541 gccgtcgaac cactccgggg ccatgatgtt gggcaggctg ccgatcagcg gctggatcag
     3601 caggccgtca gccacggctt gccgttcctc ggcgctgagg tgcgccccaa gggcgtgcag
     3661 gcggtgatga atggcggtgg gcgggtcttc ggcgggcagg tagatcaccg ggccggtggg
     3721 cagttcgccc acctccagca gatccggccc gcctgcaatc tgtgcggcca gttgcagggc
     3781 cagcatggat ttaccggcac caccgggcga caccagcgcc ccgaccgtac cggccaccat
     3841 gttgggcaaa acgtagtcca gcggtggcgg cgctgctgcg aacgcctcca gaatattgat
     3901 aggcttatgg gtagccattg attgcctcct ttgcaggcag ttggtggtta ggcgctggcg
     3961 gggtcactac ccccgccctg cgccgctctg agttcttcca ggcactcgcg cagcgcctcg
     4021 tattcgtcgt cggtcagcca gaacttgcgc tgacgcatcc ctttggcctt catgcgctcg
     4081 gcatatcgcg cttggcgtac agcgtcaggg ctggccagca ggtcgccggt ctgcttgtcc
     4141 ttttggtctt tcatatcagt caccgagaaa cttgccgggg ccgaaaggct tgtcttcgcg
     4201 gaacaaggac aaggtgcagc cgtcaaggtt aaggctggcc atatcagcga ctgaaaagcg
     4261 gccagcctcg gccttgtttg acgtataacc aaagccaccg ggcaaccaat agcccttgtc
     4321 acttttgatc aggtagaccg accctgaagc gcttttttcg tattccataa aacccccttc
     4381 tgtgcgtgag tactcatagt ataacaggcg tgagtaccaa cgcaagcact acatgctgaa
     4441 atctggcccg cccctgtcca tgcctcgctg gcggggtgcc ggtgcccgtg ccagctcggc
     4501 ccgcgcaagc tggacgctgg gcagacccat gaccttgctg acggtgcgct cgatgtaatc
     4561 cgcttcgtgg ccgggcttgc gctctgccag cgctgggctg gcctcggcca tggccttgcc
     4621 gatttcctcg gcactgcggc cccggctggc cagcttctgc gcggcgataa agtcgcactt
     4681 gctgaggtca tcaccgaagc gcttgaccag cccggccatc tcgctgcggt actcgtccag
     4741 cgccgtgcgc cggtggcggc taagctgccg ctcgggcagt tcgaggctgg ccagcctgcg
     4801 ggccttctcc tgctgccgct gggcctgctc gatctgctgg ccagcctgct gcaccagcgc
     4861 cgggccagcg gtggcggtct tgcccttgga ttcacgcagc agcacccacg gctgataacc
     4921 ggcgcgggtg gtgtgcttgt ccttgcggtt ggtgaagccc gccaagcggc catagtggcg
     4981 gctgtcggcg ctggccgggt cggcgtcgta ctcgctggcc agcgtccggg caatctgccc
     5041 ccgaagttca ccgcctgcgg cgtcggccac cttgacccat gcctgatagt tcttcgggct
     5101 ggtttccact accagggcag gctcccggcc ctcggctttc atgtcatcca ggtcaaactc
     5161 gctgaggtcg tccaccagca ccagaccatg ccgctcctgc tcggcgggcc tgatatacac
     5221 gtcattgccc tgggcattca tccgcttgag ccatggcgtg ttctggagca cttcggcggc
     5281 tgaccattcc cggttcatca tctggccggt ggtggcgtcc ctgacgccga tatcgaagcg
     5341 ctcacagccc atggccttga gctgtcggcc tatggcctgc aaagtcctgt cgttcttcat
     5401 cgggccacca agcgattccc acacattata cgagccggaa gcataaagtg taaagcctag
     5461 atccgaagga tgagccgggc tgaatgatcg accgagacag gccctgcggg gctgcacacg
     5521 cgcccccacc cttcgggtag ggggaaaggc cgctaaagcg gctaaaagcg ctccagcgta
     5581 tttctgcggg gtttggtgtg gggtttagcg ggctttgccc gcctttcccc ctgccgcgca
     5641 gcggtggggc ggtgtgtagc ctagcgcagc gaatagacca gctatccggc ctctggccgg
     5701 gcatattggg caagggcagc agcgccccac aagggcgctg ataaccgcgc ctagtggatt
     5761 attcttagat aatcatggat ggatttttcc aacaccccgc cagcccccgc ccctgctggg
     5821 tttgcaggtt tgggggcgtg acagttattg caggggttcg tgacagttat tgcagggggg
     5881 cgtgacagtt attgcagggg ttcgtgacag ttagggcgcg cccagctgtc tagggcggcg
     5941 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     6001 ctttcgactg agcctttcgt tttatttgat gcctttaatt aatagctttc gctaaggatg
     6061 atttctg
//
