LOCUS       sequence-9597-ex        7541 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 7541)
  AUTHORS   Hackett EA, Esch RK, Maleri S, Errede B
  TITLE     A family of destabilized cyan fluorescent proteins as 
            transcriptional reporters in S. cerevisiae.
  JOURNAL   Yeast. 2006 Apr 15. 23(5):333-49.
  PUBMED    16598699
REFERENCE   2  (bases 1 to 7541)
  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. 2006
            Apr 15. 23(5):333-49."
COMMENT     Sequence Label: pNC843
FEATURES             Location/Qualifiers
     source          1..7541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        866..1307
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    866..983
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1278..1301
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1710..1937
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1950..2660
                     /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
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1977..2005)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2600..2620
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2684..2871
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2739..2763)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2946..4146
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        2946..3289
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3290..3943
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      3949..4146
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4670..4687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4670..4686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4679..4701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4899..5354
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4986..5005)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5196..5217
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5371..5390)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5490..5512
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5550..5568)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5636..5740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5741..5809,5810..6601)
                     /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     complement(5959..5978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6772..7360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7261..7280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7514..7531
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaattcgagc
      241 tctggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
      301 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
      361 acacccggtg tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg
      421 gatgatgtgg tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag
      481 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
      541 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
      601 tcacaaatta gagcttcaat ttaattatat cagttattac ccggtacccc cctcgaggaa
      661 ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac
      721 atggcattac caccatatac atatccatat ctaatcttac ttatatgttg tggaaatgta
      781 aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag taatacgctt
      841 aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggcg acagccctcc
      901 gacggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat
      961 gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta
     1021 tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaattaac gaatcaaatt
     1081 aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca
     1141 gcgaagcgat gatttttgat ctattaacag atatataaat ggaaaagctg cataaccact
     1201 ttaactaata ctttcaacat tttcagtttg tattacttct tattcaaatg tcataaaagt
     1261 atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa aaaccccgga
     1321 tctgccaggc gtgtatatag cgtggatggc caggcaactt tagtgctgac acatacaggc
     1381 atatatatat gtgtgcgacg acacatgatc atatggcatg catgtgctct gtatgtatat
     1441 aaaactcttg ttttcttctt ttctctaaat attctttcct tatacattag gtcctttgta
     1501 gcataaatta ctatacttct atagacacgc aaacacaaat acacacacta aattaccgga
     1561 tcaattcgcc tgcttatctt tcttcttccg aaagtgctac ttcagaaaga gcaagaactg
     1621 tacggataag gataagtata tcttctatct actacaatac gaaaagaact ctcgaactct
     1681 ccctcccact ttactttaac taatagatta tgcagatttt cgtcaagact ttgaccggta
     1741 aaaccataac attggaagtt gaatcttccg ataccatcga caacgttaag tcgaaaattc
     1801 aagacaagga aggtatccct ccagatcaac aaagattgat ctttgccggt aagcagctag
     1861 aagacggtag aacgctgtct gattacaaca ttcagaagga gtccacctta catcttgtgc
     1921 taaggctaag aggtggtatg cacggatcca gtaaaggaga agaacttttc actggagttg
     1981 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
     2041 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
     2101 aactacctgt tccatggcca acacttgtca ctactttgac ttggggtgtt caatgctttt
     2161 caagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt
     2221 atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag
     2281 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag
     2341 aagatggaaa cattcttgga cacaaattgg aatacaacta tatttcacac aatgtataca
     2401 tcactgcaga caaacaaaag aatggaatca aagctcattt caaaattaga cacaacattg
     2461 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     2521 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     2581 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     2641 gcatggatga actatacaaa tagggcgcgc cacttctaaa taagcgaatt tcttatgatt
     2701 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2761 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2821 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     2881 tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     2941 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     3001 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     3061 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     3121 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     3181 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     3241 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgggtaggag
     3301 ggcttttgta gaaagaaata cgaacgaaac gaaaatcagc gttgccatcg ctttggacaa
     3361 agctccctta cctgaagagt cgaattttat tgatgaactt ataacttcca agcatacaaa
     3421 ccaaaaggga gaacaagtaa tccaagtaga cacgggaatt ggattcttgg atcacatgta
     3481 tcatgcactg gctaaacatg caggctggag cttacgactt tactcaagag gtgatttaat
     3541 catcgatgat catcacactg cagaagatac tgctattgca cttggtattg cattcaagca
     3601 ggctatgagt aactttgccg gcgttaaaag atttggacat gcttattgtc cacttgacga
     3661 agctctttct agaagcgtag ttgacttgtc gggacggccc tatgctgtta tcgatttggg
     3721 attaaagcgt gaaaaggttg gggaattgtc ctgtgaaatg atccctcact tactatattc
     3781 cttttcggta gcagctggaa ttactttgca tgttacctgc ttatatggta gtaatgacca
     3841 tcatcgtgct gaaagcgctt ttaaatctct ggctgttgcc atgcgcgcgg ctactagtct
     3901 tactggaagt tctgaagtcc caagcacgaa gggagtgttg taaagagtac tgacaataaa
     3961 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     4021 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     4081 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     4141 atactgctgt cgattcgata ctaacgctcg acctgcaggc atgaagatgg gaatctcggt
     4201 cgtaatgatt tctataatga cgaaaaaaaa aaaattggaa agaaaaagct tcatggcctt
     4261 tataaaaagg aactatccaa tacctcgcca gaaccaagta acagtatttt acggggcaca
     4321 aatcaagaac aataagacag gactgtaaag atggacgcat tgaactccaa agaacaacaa
     4381 gagttccaaa aagtagtgga acaaaagcaa atgaaggatt tcatgcgttt gtactctaat
     4441 ctggtagaaa gatgtttcac agactgtgtc aatgacttca caacatcaaa gctaaccaat
     4501 aaggaacaaa catgcatcat gaagtgctca gaaaagttct tgaagcatag cgaacgtgta
     4561 gggcagcgtt tccaagaaca aaacgctgcc ttgggacaag gcttgggccg ataaggtgta
     4621 ctggcgtata tatatctaat tatgtatctc tggtgtcgac aagcttggca ctggccgtcg
     4681 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4741 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4801 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     4861 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4921 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4981 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5041 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5101 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5161 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5221 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     5281 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5341 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5401 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5461 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5521 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5581 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5641 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5701 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5761 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5821 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5881 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5941 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6001 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6061 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6121 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6181 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6241 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6301 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6361 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6421 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6481 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6541 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6601 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6661 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6721 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6781 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6841 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6901 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6961 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7021 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7081 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7141 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7201 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7261 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7321 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7381 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7441 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7501 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
