LOCUS       sequence-61660-e        4114 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 4114)
  TITLE     Formosa lab yeast 3' tagging constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4114)
  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            "Unpublished"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: pTF257 (pFA6a-TEV-6xGly-His3MX)
FEATURES             Location/Qualifiers
     source          1..4114
                     /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..748
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      807..994
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(862..886)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1069..1412
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1413..2111
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2115..2312
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2416..2435)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2417..2435)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2522..2539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2693..3281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2773..2792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3452..4114,1..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     4075..4094
                     /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 aattaacgag aatttatact tccaaggttc tggtggaggc ggtggaggct ctagaggtac
      781 cctcgaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
      841 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
      901 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
      961 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     1021 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     1081 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     1141 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     1201 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
     1261 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
     1321 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
     1381 agttctcaca tcacatccga acataaacaa ccatggcaga accagcccaa aaaaagcaaa
     1441 aacaaactgt tcaggagcgc aaggcgttta tctcccgtat cactaatgaa actaaaattc
     1501 aaatcgctat ttcgctgaat ggtggttata ttcaaataaa agattcgatt cttcctgcaa
     1561 agaaggatga cgatgtagct tcccaagcta ctcagtcaca ggtcatcgat attcacacag
     1621 gtgttggctt tttggatcat atgatccatg cgttggcaaa acactctggt tggtctctta
     1681 ttgttgaatg tatcggtgac ctgcacattg acgatcacca tactaccgaa gattgcggta
     1741 tcgcattagg gcaagcgttc aaagaagcaa tgggtgctgt ccgtggtgta aaaagattcg
     1801 gtactgggtt cgcaccattg gatgaggcgc tatcacgtgc cgtagtcgat ttatctaata
     1861 gaccatttgc tgtaatcgac cttggattga agagagagat gattggtgat ttatccactg
     1921 aaatgattcc acactttttg gaaagtttcg cggaggcggc cagaattact ttgcatgttg
     1981 attgtctgag aggtttcaac gatcaccaca gaagtgagag tgcgttcaag gctttggctg
     2041 ttgccataag agaagctatt tctagcaatg gcaccaatga cgttccctca accaaaggtg
     2101 ttttgatgtg aagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat
     2161 agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt
     2221 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     2281 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc
     2341 cagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg
     2401 cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc
     2461 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2881 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3121 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3361 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3421 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3481 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3541 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3601 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3661 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3721 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3781 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3841 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3901 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3961 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4021 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4081 ccgcgccaca tagcagaact ttaaaagtgc tcat
//
