LOCUS       sequence-9604-ex        4454 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 4454)
  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 4454)
  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: pNC866
FEATURES             Location/Qualifiers
     source          1..4454
                     /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        466..907
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    466..583
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     878..901
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1310..1537
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     primer_bind     complement(1583..1600)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1583..1599)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1592..1614)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1812..2267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1899..1918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2109..2130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2284..2303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2403..2425
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2463..2481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2549..2653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2654..2722,2723..3514)
                     /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(2872..2891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3685..4273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4174..4193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4427..4444
                     /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 tcggtacccc cctcgaggaa ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag
      301 aagtttaata atcatattac atggcattac caccatatac atatccatat ctaatcttac
      361 ttatatgttg tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg
      421 gaactttcag taatacgctt aactgctcat tgctatattg aagtacggat tagaagccgc
      481 cgagcgggcg acagccctcc gacggaagac tctcctccgt gcgtcctcgt cttcaccggt
      541 cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac
      601 aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct
      661 tcaaattaac gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt
      721 atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat
      781 ggaaaagctg cataaccact ttaactaata ctttcaacat tttcagtttg tattacttct
      841 tattcaaatg tcataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt
      901 caaggagaaa aaaccccgga tctgccaggc gtgtatatag cgtggatggc caggcaactt
      961 tagtgctgac acatacaggc atatatatat gtgtgcgacg acacatgatc atatggcatg
     1021 catgtgctct gtatgtatat aaaactcttg ttttcttctt ttctctaaat attctttcct
     1081 tatacattag gtcctttgta gcataaatta ctatacttct atagacacgc aaacacaaat
     1141 acacacacta aattaccgga tcaattcgcc tgcttatctt tcttcttccg aaagtgctac
     1201 ttcagaaaga gcaagaactg tacggataag gataagtata tcttctatct actacaatac
     1261 gaaaagaact ctcgaactct ccctcccact ttactttaac taatagatta tgcagatttt
     1321 cgtcaagact ttgaccggta aaaccataac attggaagtt gaatcttccg ataccatcga
     1381 caacgttaag tcgaaaattc aagacaagga aggtatccct ccagatcaac aaagattgat
     1441 ctttgccggt aagcagctag aagacggtag aacgctgtct gattacaaca ttcagaagga
     1501 gtccacctta catcttgtgc taaggctaag aggtggtgaa cacggatcct ctagagtcga
     1561 cctgcaggca tgcaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     1621 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     1681 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     1741 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     1801 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1861 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1921 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1981 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     2041 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2101 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     2161 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2221 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     2281 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2341 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2401 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2461 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2521 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2581 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2641 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2701 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2761 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2821 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2881 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2941 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3001 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3061 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3121 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3181 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3241 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3301 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3361 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3421 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3481 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3541 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3601 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3661 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3721 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3781 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3841 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3901 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3961 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4021 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4081 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4141 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4201 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4261 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4321 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4381 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4441 cgaggaagcg gaag
//
