LOCUS       sequence-61673-e        4260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4260)
  TITLE     Formosa lab yeast 3' tagging constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4260)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTF269 (pFA6a-PSc-6xGly-12xHis-KanMX)
FEATURES             Location/Qualifiers
     source          1..4260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(199..303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     371..389
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(427..449)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     549..568
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        649..667
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     649..666
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             728..751
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             776..805
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             779..808
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             782..811
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     terminator      840..1027
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(895..919)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1102..2458
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1102..1445
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1446..2255
                     /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(1513..1532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2261..2458
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2562..2581)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2563..2581)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2668..2685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2839..3427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2919..2938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3598..4260,1..129,130..198))
                     /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     4221..4240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
       61 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
      421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
      481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      541 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
      601 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
      661 ctatagaacg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt
      721 aattaacttg gaagtattgt tccaaggtcc aggtggaggc ggtggaggct ctagacatca
      781 ccaccatcat caccaccatc atcatcacca ttgactcgag gcgcgccact tctaaataag
      841 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
      901 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
      961 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     1021 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
     1081 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     1141 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     1201 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     1261 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     1321 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     1381 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     1441 caaccatggg taaggaaaag actcacgttt cgaggccgcg attaaattcc aacatggatg
     1501 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1561 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1621 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1681 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1741 tccccggcaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1801 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1861 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1921 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1981 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2041 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2101 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2161 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2221 tgcagtttca tttgatgctc gatgagtttt tctaatcagt actgacaata aaaagattct
     2281 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     2341 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     2401 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     2461 gtcgattcga tactaacgcc gccatccagt ttaaacgagc tcgaattcat cgatgatatc
     2521 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     2581 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3001 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     3061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3241 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
//
