LOCUS       sequence-455513-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSEVAb (SEVA 3.1), Type IIS compatible plasmid with sfGFP under the 
            BBa_J23119 promoter, pSEVAb82 backbone, RK2 ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  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 4520)
  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: pSEVAb827
FEATURES             Location/Qualifiers
     source          1..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(67..153)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    193..214
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        215..249
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     RBS             258..269
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    996..1016
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1042..1136
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1243..2043
                     /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            2219..2327
                     /note="incP origin of transfer"
     CDS             complement(2388..3536)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      3929..4460
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 aaaaaaagga tctcaagaag atcctttgat cttttctacg gcgcgcccag ctgtctaggg
       61 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      121 ccagtctttc gactgagcct ttcgttttat ttgatgcctt taattaatag ctttcgctaa
      181 ggatgatttc tggaattcgc ggccgcttct agagttgacg gctagctcag tcctaggtat
      241 tgtgctagct actagagaaa gaggagaaat actagatgcg taaaggcgaa gaactgttca
      301 cgggcgtagt tccgattctg gtcgagctgg acggcgatgt gaacggtcat aagtttagcg
      361 ttcgcggtga aggtgagggc gacgcgacca acggcaaact gaccctgaag ttcatctgca
      421 ccaccggtaa actgccggtg ccttggccga ccttggtgac gacgttgacg tatggcgtgc
      481 agtgttttgc gcgttatccg gaccacatga aacaacacga tttcttcaaa tctgcgatgc
      541 cggagggtta cgtccaggag cgtaccattt ccttcaagga tgatggctac tacaaaactc
      601 gcgcagaggt taagtttgaa ggtgacacgc tggtcaatcg tatcgaattg aagggtatcg
      661 actttaaaga ggatggtaac attctgggcc ataaactgga gtataacttc aacagccata
      721 atgtttacat tacggcagac aagcaaaaga acggcatcaa ggccaatttc aagattcgcc
      781 acaatgttga ggacggtagc gtccaactgg ccgaccatta ccagcagaac accccaattg
      841 gtgacggtcc ggttttgctg ccggataatc actatctgag cacccaaagc gtgctgagca
      901 aagatccgaa cgaaaaacgt gatcacatgg tcctgctgga atttgtgacc gctgcgggca
      961 tcacccacgg tatggatgaa ctgtacaaat aataatacta gtagcggccg ctgcagtccg
     1021 gcaaaaaagg gcaaggtctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     1081 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag
     1141 gggtccccaa taattacgat ttaaatttgt gtctcaaaat ctctgatgtt acattgcaca
     1201 agataaaaat atatcatcat gaacaataaa actgtctgga atatgtcatc agcggtggag
     1261 tgcaatgtcg tgcaatacga atggcgaaaa gccgagctca tcggtcagct tctcaacctt
     1321 ggggttaccc ccggcggtgt gctgctggtc cacagctcct tccgtagcgt ccggcccctc
     1381 gaagatgggc cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc gggagggacg
     1441 ctcgtcatgc cctcgtggtc aggtctggac gacgagccgt tcgatcctgc cacgtcgccc
     1501 gttacaccgg accttggagt tgtctctgac acattctggc gcctgccaaa tgtaaagcgc
     1561 agcgcccatc catttgcctt tgcggcagcg gggccacagg cagagcagat catctctgat
     1621 ccattgcccc tgccacctca ctcgcctgca agcccggtcg cccgtgtcca tgaactcgat
     1681 gggcaggtac ttctcctcgg cgtgggacac gatgccaaca cgacgctgca tcttgccgag
     1741 ttgatggcaa aggttcccta tggggtgccg agacactgca ccattcttca ggatggcaag
     1801 ttggtacgcg tcgattatct cgagaatgac cactgctgtg agcgctttgc cttggcggac
     1861 aggtggctca aggagaagag ccttcagaag gaaggtccag tcggtcatgc ctttgctcgg
     1921 ttgatccgct cccgcgacat tgtggcgaca gccctgggtc aactgggccg agatccgttg
     1981 atcttcctgc atccgccaga ggcgggatgc gaagaatgcg atgccgctcg ccagtcgatt
     2041 ggctgagctc atagtccaat tggaccgcgg tccgcgcgtt gtccttttcc gctgcataac
     2101 cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata
     2161 caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg
     2221 gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt
     2281 cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggccgcg
     2341 atgcaggtgg ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa
     2401 tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg
     2461 caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg
     2521 aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac
     2581 atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg
     2641 tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc
     2701 ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg
     2761 cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc
     2821 gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag
     2881 gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt
     2941 tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg
     3001 tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac
     3061 agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata
     3121 tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc
     3181 tgcttggctt cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc
     3241 gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttcg
     3301 agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc
     3361 acgctgtcgc gctcgatctt ggccgtagct tgctggacta tcgagccgac ggactggaag
     3421 gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa
     3481 aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac
     3541 cctccttgcg ggattgcccc ggaattaatt ccccggatcg atccgtcgat cttgatcccc
     3601 tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa
     3661 ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc
     3721 agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt
     3781 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     3841 gactggcttt ctacgtggct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca
     3901 gcccctgggg ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca
     3961 ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt
     4021 ataaatattg gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg
     4081 aaacccttgc aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc
     4141 gcccctcatc tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca
     4201 gtagtcgcgc ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc
     4261 atctgtggga aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc
     4321 cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg
     4381 gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg
     4441 tatccacaac gccggcggcc ctacatggct ctgctgtagt gagtgggttg cgctccggca
     4501 gcggtcctga tcccccgcag
//
