LOCUS       sequence-384247-        3988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA6a derived selection cassette 5' flanked with tDEG1 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 3988)
  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 3988)
  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: kanMX6-ins6
FEATURES             Location/Qualifiers
     source          1..3988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            177..1533
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        177..520
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             521..1330
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(588..607)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1336..1533
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1637..1656)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1638..1656)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1743..1760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1914..2502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1994..2013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2673..3464,3465..3533))
                     /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     3296..3315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3534..3638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3706..3724
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3762..3784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3884..3903
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3984..3988,1..14)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3984..3988,1..13)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ggtgacacta tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcc
       61 ccgggttaat taagaaaaga aatatagtct tcaaggttat attatacagg tttatatatt
      121 attggcgcgc cagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca
      181 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
      241 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
      301 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg
      361 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
      421 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
      481 taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca
      541 cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
      601 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
      661 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
      721 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
      781 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt
      841 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
      901 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
      961 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1021 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc
     1081 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     1141 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     1201 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     1261 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     1321 gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     1381 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     1441 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     1501 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     1561 ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc
     1621 gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg
     1681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3661 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3721 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3781 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3841 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     3901 gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata
     3961 accttatgta tcatacacat acgattta
//
