LOCUS       sequence-128050-        5546 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 5546)
  AUTHORS   Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat 
            A, Philippsen P, Pringle JR
  TITLE     Additional modules for versatile and economical PCR-based gene 
            deletion and modification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 1998 Jul;14(10):953-61.
  PUBMED    9717241
REFERENCE   2  (bases 1 to 5546)
  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: "Yeast."; 
            date: "1998-07"; volume: "14(10)"; pages: "953-61"
COMMENT     Sequence Label: pFA6a-GFP(S65T)-His3MX6
FEATURES             Location/Qualifiers
     source          1..5546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             61..771
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(88..116)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     711..731
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      795..982
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(850..874)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1057..1400
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1401..2099
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2103..2300
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2404..2423)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2405..2423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2510..2527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2681..3269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2761..2780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3440..4231,4232..4300))
                     /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     4063..4082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4301..4405)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  4466..5233
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     primer_bind     5250..5268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5306..5328)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5428..5447
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5528..5546
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5528..5545
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
      121 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
      181 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
      241 actactttca cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat
      301 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa
      361 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
      421 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
      481 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
      541 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
      601 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
      661 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
      721 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagggcgcg
      781 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
      841 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
      901 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
      961 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
     1021 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     1081 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     1141 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     1201 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     1261 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     1321 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     1381 acatccgaac ataaacaacc atggcagaac cagcccaaaa aaagcaaaaa caaactgttc
     1441 aggagcgcaa ggcgtttatc tcccgtatca ctaatgaaac taaaattcaa atcgctattt
     1501 cgctgaatgg tggttatatt caaataaaag attcgattct tcctgcaaag aaggatgacg
     1561 atgtagcttc ccaagctact cagtcacagg tcatcgatat tcacacaggt gttggctttt
     1621 tggatcatat gatccatgcg ttggcaaaac actctggttg gtctcttatt gttgaatgta
     1681 ttggtgacct gcacattgac gatcaccata ctaccgaaga ttgcggtatc gcattagggc
     1741 aagcgttcaa agaagcaatg ggtgctgtcc gtggtgtaaa aagattcggt actgggttcg
     1801 caccattgga tgaggcgcta tcacgtgccg tagtcgattt atctagtaga ccatttgctg
     1861 taatcgacct tggattgaag agagagatga ttggtgattt atccactgaa atgattccac
     1921 actttttgga aagtttcgcg gaggcggcca gaattacttt gcatgttgat tgtctgagag
     1981 gtttcaacga tcaccacaga agtgagagtg cgttcaaggc tttggctgtt gccataagag
     2041 aagctatttc tagcaatggc accaatgacg ttccctcaac caaaggtgtt ttgatgtgaa
     2101 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     2161 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     2221 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     2281 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga
     2341 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg
     2401 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
     2461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4441 aaaccattat tatcatgaca ttaacggtaa tgactccaac ttattgatag tgttttatgt
     4501 tcagataatg cccgatgact ttgtcatgca gctccaccga ttttgagaac gacagcgact
     4561 tccgtcccag ccgtgccagg tgctgcctca gattcaggtt atgccgctca attcgctgcg
     4621 tatatcgctt gctgattacg tgcagctttc ccttcaggcg ggattcatac agcggccagc
     4681 catccgtcat ccatatcacc acgtcaaagg gtgacagcag gctcataaga cgccccagcg
     4741 tcgccatagt gcgttcaccg aatacgtgcg caacaaccgt cttccggaga ctgtcatacg
     4801 cgtaaaacag ccagcgctgg cgcgatttag ccccgacata gccccactgt tcgtccattt
     4861 ccgcgcagac gatgacgtca ctgcccggct gtatgcgcga ggttaccgac tgcggcctga
     4921 gttttttaag tgacgtaaaa tcgtgttgag gccaacgccc ataatgcggg ctgttgcccg
     4981 gcatccaacg ccattcatgg ccatatcaat gattttctgg tgcgtaccgg gttgagaagc
     5041 ggtgtaagtg aactgcagtt gccatgtttt acggcagtga gagcagagat agcgctgatg
     5101 tccggcggtg cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct
     5161 gatagaaaca gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt
     5221 tggcagcatc accgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5281 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5341 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5401 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5461 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     5521 acatacgatt taggtgacac tataga
//
