LOCUS       sequence-61686-e        4522 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 4522)
  TITLE     Formosa lab yeast 3' tagging constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4522)
  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            "Unpublished"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: pZC09 (pFA6a-TEV-6xGly-PrtA-His3MX)
FEATURES             Location/Qualifiers
     source          1..4522
                     /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"
     CDS             809..982
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             983..1156
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     terminator      1215..1402
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1270..1294)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1477..1820
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1821..2519
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2523..2720
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2824..2843)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2825..2843)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2930..2947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3101..3689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3181..3200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3860..4522,1..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     4483..4502
                     /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 ctagaaaaac
      781 cgcggctctt gcgcaacacg atgaagccgt ggacaacaaa ttcaacaaag aacaacaaaa
      841 cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccttcat
      901 ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct
      961 aaatgatgct caggcgccga aagtagacaa caaattcaac aaagaacaac aaaacgcgtt
     1021 ctatgagatc ttacatttac ctaacttaaa cgaagaacaa cgaaacgcct tcatccaaag
     1081 tttaaaagat gacccaagcc aaagcgctaa ccttttagca gaagctaaaa agctaaatgg
     1141 tgctcaggcg ccgaaagtag acgcgaattc cgcggggaag tcaacctgac tcgaggcgcg
     1201 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     1261 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     1321 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
     1381 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
     1441 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     1501 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     1561 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     1621 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     1681 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     1741 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     1801 acatccgaac ataaacaacc atggcagaac cagcccaaaa aaagcaaaaa caaactgttc
     1861 aggagcgcaa ggcgtttatc tcccgtatca ctaatgaaac taaaattcaa atcgctattt
     1921 cgctgaatgg tggttatatt caaataaaag attcgattct tcctgcaaag aaggatgacg
     1981 atgtagcttc ccaagctact cagtcacagg tcatcgatat tcacacaggt gttggctttt
     2041 tggatcatat gatccatgcg ttggcaaaac actctggttg gtctcttatt gttgaatgta
     2101 tcggtgacct gcacattgac gatcaccata ctaccgaaga ttgcggtatc gcattagggc
     2161 aagcgttcaa agaagcaatg ggtgctgtcc gtggtgtaaa aagattcggt actgggttcg
     2221 caccattgga tgaggcgcta tcacgtgccg tagtcgattt atctaataga ccatttgctg
     2281 taatcgacct tggattgaag agagagatga ttggtgattt atccactgaa atgattccac
     2341 actttttgga aagtttcgcg gaggcggcca gaattacttt gcatgttgat tgtctgagag
     2401 gtttcaacga tcaccacaga agtgagagtg cgttcaaggc tttggctgtt gccataagag
     2461 aagctatttc tagcaatggc accaatgacg ttccctcaac caaaggtgtt ttgatgtgaa
     2521 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     2581 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     2641 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     2701 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga
     2761 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg
     2821 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
     2881 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     3301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3541 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3901 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3961 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4021 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4081 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4141 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4201 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4261 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4321 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4381 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4441 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4501 gcagaacttt aaaagtgctc at
//
