LOCUS       sequence-6754-ex        5578 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 5578)
  AUTHORS   Sheff MA, Thorn KS
  TITLE     Optimized cassettes for fluorescent protein tagging in Saccharomyces
            cerevisiae.
  JOURNAL   Yeast 2004 Jun;21(8):661-70.
  PUBMED    15197731
REFERENCE   2  (bases 1 to 5578)
  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: "2004-06"; volume: "21(8)"; pages: "661-70"
COMMENT     Sequence Label: pKT0176
FEATURES             Location/Qualifiers
     source          1..5578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..687
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             730..1404
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     terminator      1425..1612
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1480..1504)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1687..2030
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2849..3046
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3153..3172)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3154..3172)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3259..3276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3430..4018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3510..3529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4189..4980,4981..5049))
                     /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     4812..4831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5050..5154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5222..5240
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5278..5300)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5400..5419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5500..5518
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5500..5517
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttaattaaca tggtggcctc ctccgaggac gtcatcaaag agttcatgcg cttcaaggtg
       61 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      121 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      181 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggcgtacgt gaagcacccc
      241 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      301 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      361 acgctgatct acaaggtgaa gttccgcggc accaacttcc cccccgacgg ccccgtaatg
      421 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
      481 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag
      541 ttcaagacca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac
      601 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
      661 tccgagggcc gccaccacct gttcctgggg catggcaccg gcagcaccgg cagcggcagc
      721 tccggcaccg cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg
      781 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      841 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      901 gacatcctgt ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac
      961 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1021 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     1081 atctacaagg tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1141 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1201 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1261 accatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag
     1321 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     1381 ggccgccacc acctgttcct gtagggcgcg ccacttctaa ataagcgaat ttcttatgat
     1441 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     1501 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     1561 tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc ggcagatccg
     1621 ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc gggtcacccg
     1681 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     1741 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     1801 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
     1861 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
     1921 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
     1981 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgacagtca
     2041 acactaagac ctatagtgag agagcagaaa ctcatgcctc accagtagca cagcgattat
     2101 ttcgattaat ggaactgaag aaaaccaatt tatgtgcatc aattgacgtt gataccacta
     2161 aggaattcct tgaattaatt gataaattag gtccttatgt atgcttaatc aagactcata
     2221 ttgatataat caatgatttt tcctatgaat ccactattga accattatta gaactttcac
     2281 gtaaacatca atttatgatt tttgaagata gaaaatttgc tgatattggt aataccgtaa
     2341 agaaacaata tattggtgga gtttataaaa ttagtagttg ggcagatatt accaatgctc
     2401 atggtgtcac tgggaatgga gtggttgaag gattaaaaca gggagctaaa gaaaccacca
     2461 ccaaccaaga gccaagaggg ttattgatgt tagctgaatt atcatcagtg ggatcattag
     2521 catatggaga atattctcaa aaaactgttg aaattgctaa atccgataag gaatttgtta
     2581 ttggatttat tgcccaacgt gatatgggtg gccaagaaga aggatttgat tggcttatta
     2641 tgacacctgg agttggatta gatgataaag gtgatggatt aggacaacaa tatagaactg
     2701 ttgatgaagt tgttagcact ggaactgata ttatcattgt tggtagagga ttgtttggta
     2761 aaggaagaga tccagatatt gaaggtaaaa ggtatagaaa tgctggttgg aatgcttatt
     2821 tgaaaaagac tggccaatta taatcagtac tgacaataaa aagattcttg ttttcaagaa
     2881 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     2941 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     3001 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     3061 ctaacgccgc catccagtgt cgaaaacgag ctcgaattca tcgatgatat cagatccact
     3121 agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     3181 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3241 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3601 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     3661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3841 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4681 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4741 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4801 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4861 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4921 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4981 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5041 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5161 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5221 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5281 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5341 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5401 ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg
     5461 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaac
     5521 gcggccgcca gctgaagctt cgtacgctgc aggtcgacgg atcggtgacg gtgctggt
//
