LOCUS       sequence-444281-        3683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSEVAb (SEVA 3.1), Type IIS compatible plasmid with sfGFP under the 
            BBa_J23119 promoter, pSEVAb89 backbone, pBR322/ROP ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3683)
  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 3683)
  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: pSEVAb897
FEATURES             Location/Qualifiers
     source          1..3683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    733..873
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(799..818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     959..981
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(975..1166)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(1346..1432)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1472..1493
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        1494..1528
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     RBS             1537..1548
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    2275..2295
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2321..2415
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2522..3322
                     /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            3498..3606
                     /note="incP origin of transfer"
     rep_origin      join(3642..3683,1..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 actgtccttc 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 caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      781 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      841 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      901 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      961 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     1021 taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc atccgcgtcc
     1081 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     1141 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     1201 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     1261 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcggggg
     1321 cgcgcccagc tgtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac
     1381 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgccttt
     1441 aattaatagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagttgacgg
     1501 ctagctcagt cctaggtatt gtgctagcta ctagagaaag aggagaaata ctagatgcgt
     1561 aaaggcgaag aactgttcac gggcgtagtt ccgattctgg tcgagctgga cggcgatgtg
     1621 aacggtcata agtttagcgt tcgcggtgaa ggtgagggcg acgcgaccaa cggcaaactg
     1681 accctgaagt tcatctgcac caccggtaaa ctgccggtgc cttggccgac cttggtgacg
     1741 acgttgacgt atggcgtgca gtgttttgcg cgttatccgg accacatgaa acaacacgat
     1801 ttcttcaaat ctgcgatgcc ggagggttac gtccaggagc gtaccatttc cttcaaggat
     1861 gatggctact acaaaactcg cgcagaggtt aagtttgaag gtgacacgct ggtcaatcgt
     1921 atcgaattga agggtatcga ctttaaagag gatggtaaca ttctgggcca taaactggag
     1981 tataacttca acagccataa tgtttacatt acggcagaca agcaaaagaa cggcatcaag
     2041 gccaatttca agattcgcca caatgttgag gacggtagcg tccaactggc cgaccattac
     2101 cagcagaaca ccccaattgg tgacggtccg gttttgctgc cggataatca ctatctgagc
     2161 acccaaagcg tgctgagcaa agatccgaac gaaaaacgtg atcacatggt cctgctggaa
     2221 tttgtgaccg ctgcgggcat cacccacggt atggatgaac tgtacaaata ataatactag
     2281 tagcggccgc tgcagtccgg caaaaaaggg caaggtcttg gactcctgtt gatagatcca
     2341 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     2401 tttattggtg agaatccagg ggtccccaat aattacgatt taaatttgtg tctcaaaatc
     2461 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctggaa
     2521 tatgtcatca gcggtggagt gcaatgtcgt gcaatacgaa tggcgaaaag ccgagctcat
     2581 cggtcagctt ctcaaccttg gggttacccc cggcggtgtg ctgctggtcc acagctcctt
     2641 ccgtagcgtc cggcccctcg aagatgggcc acttggactg atcgaggccc tgcgtgctgc
     2701 gctgggtccg ggagggacgc tcgtcatgcc ctcgtggtca ggtctggacg acgagccgtt
     2761 cgatcctgcc acgtcgcccg ttacaccgga ccttggagtt gtctctgaca cattctggcg
     2821 cctgccaaat gtaaagcgca gcgcccatcc atttgccttt gcggcagcgg ggccacaggc
     2881 agagcagatc atctctgatc cattgcccct gccacctcac tcgcctgcaa gcccggtcgc
     2941 ccgtgtccat gaactcgatg ggcaggtact tctcctcggc gtgggacacg atgccaacac
     3001 gacgctgcat cttgccgagt tgatggcaaa ggttccctat ggggtgccga gacactgcac
     3061 cattcttcag gatggcaagt tggtacgcgt cgattatctc gagaatgacc actgctgtga
     3121 gcgctttgcc ttggcggaca ggtggctcaa ggagaagagc cttcagaagg aaggtccagt
     3181 cggtcatgcc tttgctcggt tgatccgctc ccgcgacatt gtggcgacag ccctgggtca
     3241 actgggccga gatccgttga tcttcctgca tccgccagag gcgggatgcg aagaatgcga
     3301 tgccgctcgc cagtcgattg gctgagctca tagtccaatt ggaccgcggt ccgcgcgttg
     3361 tccttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     3421 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     3481 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     3541 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     3601 tggccgtagg ccggccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3661 cgcgtaatct gctgcttgca aac
//
