LOCUS       sequence-384248-        4073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA6a derived selection cassette 5' flanked with tCYC1 transcription
            terminator, allows genome modification without disruption of 
            insertion neighboring genes by transcription interference.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4073)
  AUTHORS   Powers EN, Chan C, Doron-Mandel E, Llacsahuanga Allcca L, Kim Kim J,
            Jovanovic M, Brar GA
  TITLE     Bidirectional promoter activity from expression cassettes can drive 
            off-target repression of neighboring gene translation.
  JOURNAL   Elife. 2022 Dec 12;11:e81086. doi: 10.7554/eLife.81086.
  PUBMED    36503721
REFERENCE   2  (bases 1 to 4073)
  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: "Elife."; 
            date: "2022-12-12"; volume: "11:e81086. doi"; pages: " 
            10.7554/eLife.81086"
COMMENT     Sequence Label: natMX-ins7
FEATURES             Location/Qualifiers
     source          1..4073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..23
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5..22
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     245..263
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1731..1750)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1732..1750)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1837..1854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2008..2596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2088..2107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2767..3558,3559..3627))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3390..3409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3628..3732)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3800..3818
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3856..3878)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3978..3997
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgatttag gtgacactat agaacgcggc cgccagctga agcttcgtac gctgcaggtc
       61 gacggatccc cgggttaatt aatataatgt tacatgcgta cacgcgtttg tacagaaaaa
      121 aaagaaaaat ttgaaatata aataacgttc ttaatactaa cataactata aaaaaataaa
      181 tagggaccta gacttcaggt tgtctaactc cttccttttc ggttagagcg gatgtggggg
      241 gagggcgtga atgtaagcgt gacataacta attacatgat atcgacaaag gaaaaggggc
      301 ctgtgaagct ctaggagaag ccctaggaga agcaaagggg aagcccaaag agggacaagc
      361 gcaggccact acagtaacaa agacaggaag cccaaaagat cgcaccttcc agtccatcgc
      421 cttcccccat tgtcatatgg cagtttatag cagtgtggca gtatggcagt gtggcaatgt
      481 gaagcagtgt gcaatacttc caaagtggat aatgtgatgc ttgcaaaaag ttgggaccac
      541 caatgctgta gagacaatgg gataccctag cgaatgaaga aaaaaaggaa aacgaaaaaa
      601 atttgtatat aaagcagagg caatttttgt agcggcatga ttagttgtca attggcaaga
      661 cggcatacat acatctattc gataatgggt accactcttg acgacacggc ttaccggtac
      721 cgcaccagtg tcccgggaga cgccgaagcc atcgaggcac tggatgggtc cttcaccacc
      781 gacactgtct tccgcgtcac tgccaccggg gacggcttca ccctgcggga ggtgccggtg
      841 gacccgccac tgaccaaggt gttccccgat gacgagtcgg atgacgaatc tgacgacggt
      901 gaggacggcg acccagactc ccggacgttc gtcgcgtacg gggacgacgg cgacctagcg
      961 ggattcgtgg tcgtctcgta ctccggctgg aaccgacggc tgaccgtcga ggacatcgaa
     1021 gtagccccag aacaccgagg gcacggagtc gggcgagcgt tgatgggtct cgcgacggag
     1081 ttcgctcgcg agcggggcgc aggtcaccta tggctagagg tcaccaacgt caacgcacca
     1141 gcgatccacg cgtaccgacg gatggggttc accctctgcg gtctggacac cgcactgtac
     1201 gacggcacag cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaaact
     1261 agcttgagaa ctccccctag ttcttagaaa ggtgtaattt gagcgcatga ataatatttc
     1321 tccactcccg atacttttaa tagggtagta aaaaatttaa ccagttcgag tgcttcctaa
     1381 caagatgaac ataaacttga tagagtctac cttttgatag ggtataaatt ttatcttcct
     1441 gtttagagat gttttctcaa tggttaatgg aaaatatatg ctatatcttg ccacgcagta
     1501 catcctttcg aagctttttt attctatttc aacacatcga taaatgacgt acataaaaat
     1561 tataaataat ttaatgaaaa aattattatt atgacttata cattaattaa ttatcagttt
     1621 attttttttt tatttttttt ctttctcagt ttaattagtt taaacgagct cgaattcatc
     1681 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     1741 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     1801 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3721 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3781 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     3841 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3901 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3961 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4021 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac aca
//
