LOCUS       sequence-444279-        4258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSEVAb (SEVA 3.1), Type IIS compatible plasmid with sfGFP under the 
            BBa_J23119 promoter, pSEVAb84 backbone, pRO1600/ColE1 ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4258)
  AUTHORS   van den Brink M, Althuis TY, Danelon C, Claassens NJ
  TITLE     MOSAIC: A Highly Efficient, One-Step Recombineering Approach to 
            Plasmid Editing and Diversification.
  JOURNAL   ACS Synth Biol. 2025 Jun 5. doi: 10.1021/acssynbio.4c00657.
  PUBMED    40471122
REFERENCE   2  (bases 1 to 4258)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2025 Jun 5. doi: 10.1021/acssynbio.4c00657."
COMMENT     Sequence Label: pSEVAb847
FEATURES             Location/Qualifiers
     source          1..4258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(601..1434)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      1448..1799
                     /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(1872..1958)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1998..2017
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     RBS             2054..2065
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    2792..2812
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2838..2932
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3039..3839
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     oriT            4015..4123
                     /note="incP origin of transfer"
     rep_origin      join(4217..4258,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggcatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccgtgaa aggcaggccg gtccgtggtg gccacggcct
      601 ctaggccaga tccagcggca tctgggttag tcgagcgcgg gccgcttccc atgtctcacc
      661 agggcgagcc tgtttcgcga tctcagcatc tgaaatcttc ccggccttgc gcttcgctgg
      721 ggccttaccc accgccttgg cgggcttctt cggtccaaaa ctgaacaaca gatgtgtgac
      781 cttgcgcccg gtctttcgct gcgcccactc cacctgtagc gggctgtgct cgttgatctg
      841 cgtcacggct ggatcaagca ctcgcaactt gaagtccttg atcgagggat accggccttc
      901 cagttgaaac cactttcgca gctggtcaat ttctatttcg cgctggccga tgctgtccca
      961 ttgcatgagc agctcgtaaa gcctgatcgc gtgggtgctg tccatcttgg ccacgtcagc
     1021 caaggcgtat ttggtgaact gtttggtgag ttccgtcagg tacggcagca tgtctttggt
     1081 gaacctgagt tctacacggc cctcaccctc ccggtagatg attgtttgca cccagccggt
     1141 aatcatcaca ctcggtcttt tccccttgcc attgggctct tgggttaacc ggacttcccg
     1201 ccgtttcagg cgcagggccg cttctttgag ctggttgtag gaagattcga tagggacacc
     1261 cgccatcgtc gctatgtcct ccgccgtcac tgaatacatc acttcatcgg tgacaggctc
     1321 gctcctcttc acctggctaa tacaggccag aacgatccgc tgttcctgaa cactgaggcg
     1381 atacgcggcc tcgaccaggg cattgctttt gtaaaccatt gggggtgagg ccacgttcga
     1441 cattccttgt gtataagggg acactgtatc tgcgtcccac aatacaacaa atccgtccct
     1501 ttacaacaac aaatccgtcc cttcttaaca acaaatccgt cccttaatgg caacaaatcc
     1561 gtcccttttt aaactctaca ggccacggat tacgtggcct gtagacgtcc taaaaggttt
     1621 aaaagggaaa aggaagaaaa gggtggaaac gcaaaaaacg caccactacg tggccccgtt
     1681 ggggccgcat ttgtgcccct gaaggggcgg gggaggcgtc tgggcaatcc ccgttttacc
     1741 agtcccctat cgccgcctga gagggcgcag gaagcgagta atcagggtat cgaggcggat
     1801 tcacccttgg cgtccaacca gcggcaccag cggcgcctga gaggggcgcg cccagctgtc
     1861 tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
     1921 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctttaatt aatagctttc
     1981 gctaaggatg atttctggaa ttcgcggccg cttctagata gctcagtcct aggtattgtg
     2041 ctagctacta gagaaagagg agaaatacta gatgcgtaaa ggcgaagaac tgttcacggg
     2101 cgtagttccg attctggtcg agctggacgg cgatgtgaac ggtcataagt ttagcgttcg
     2161 cggtgaaggt gagggcgacg cgaccaacgg caaactgacc ctgaagttca tctgcaccac
     2221 cggtaaactg ccggtgcctt ggccgacctt ggtgacgacg ttgacgtatg gcgtgcagtg
     2281 ttttgcgcgt tatccggacc acatgaaaca acacgatttc ttcaaatctg cgatgccgga
     2341 gggttacgtc caggagcgta ccatttcctt caaggatgat ggctactaca aaactcgcgc
     2401 agaggttaag tttgaaggtg acacgctggt caatcgtatc gaattgaagg gtatcgactt
     2461 taaagaggat ggtaacattc tgggccataa actggagtat aacttcaaca gccataatgt
     2521 ttacattacg gcagacaagc aaaagaacgg catcaaggcc aatttcaaga ttcgccacaa
     2581 tgttgaggac ggtagcgtcc aactggccga ccattaccag cagaacaccc caattggtga
     2641 cggtccggtt ttgctgccgg ataatcacta tctgagcacc caaagcgtgc tgagcaaaga
     2701 tccgaacgaa aaacgtgatc acatggtcct gctggaattt gtgaccgctg cgggcatcac
     2761 ccacggtatg gatgaactgt acaaataata atactagtag cggccgctgc agtccggcaa
     2821 aaaagggcaa ggtcttggac tcctgttgat agatccagta atgacctcag aactccatct
     2881 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaggggt
     2941 ccccaataat tacgatttaa atttgtgtct caaaatctct gatgttacat tgcacaagat
     3001 aaaaatatat catcatgaac aataaaactg tctggaatat gtcatcagcg gtggagtgca
     3061 atgtcgtgca atacgaatgg cgaaaagccg agctcatcgg tcagcttctc aaccttgggg
     3121 ttacccccgg cggtgtgctg ctggtccaca gctccttccg tagcgtccgg cccctcgaag
     3181 atgggccact tggactgatc gaggccctgc gtgctgcgct gggtccggga gggacgctcg
     3241 tcatgccctc gtggtcaggt ctggacgacg agccgttcga tcctgccacg tcgcccgtta
     3301 caccggacct tggagttgtc tctgacacat tctggcgcct gccaaatgta aagcgcagcg
     3361 cccatccatt tgcctttgcg gcagcggggc cacaggcaga gcagatcatc tctgatccat
     3421 tgcccctgcc acctcactcg cctgcaagcc cggtcgcccg tgtccatgaa ctcgatgggc
     3481 aggtacttct cctcggcgtg ggacacgatg ccaacacgac gctgcatctt gccgagttga
     3541 tggcaaaggt tccctatggg gtgccgagac actgcaccat tcttcaggat ggcaagttgg
     3601 tacgcgtcga ttatctcgag aatgaccact gctgtgagcg ctttgccttg gcggacaggt
     3661 ggctcaagga gaagagcctt cagaaggaag gtccagtcgg tcatgccttt gctcggttga
     3721 tccgctcccg cgacattgtg gcgacagccc tgggtcaact gggccgagat ccgttgatct
     3781 tcctgcatcc gccagaggcg ggatgcgaag aatgcgatgc cgctcgccag tcgattggct
     3841 gagctcatag tccaattgga ccgcggtccg cgcgttgtcc ttttccgctg cataaccctg
     3901 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     3961 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     4021 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     4081 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccgataatc
     4141 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4201 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaac
//
