LOCUS       sequence-9593-ex        3839 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 3839)
  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 3839)
  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: pNC824
FEATURES             Location/Qualifiers
     source          1..3839
                     /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"
     CDS             695..922
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     primer_bind     complement(968..985)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(968..984)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(977..999)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1197..1652
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1284..1303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1494..1515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1669..1688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1788..1810
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1848..1866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1934..2038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2039..2107,2108..2899)
                     /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(2257..2276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3070..3658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3559..3578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3812..3829
                     /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 cctcgagtga tgaaacaaac atgaaacgtc tgtaatttga aacaaatcga
      301 gcagatctgc caggcgtgta tatagcgtgg atggccaggc aactttagtg ctgacacata
      361 caggcatata tatatgtgtg cgacgacaca tgatcatatg gcatgcatgt gctctgtatg
      421 tatataaaac tcttgttttc ttcttttctc taaatattct ttccttatac attaggtcct
      481 ttgtagcata aattactata cttctataga cacgcaaaca caaatacaca cactaaatta
      541 ccggatcaat tcgcctgctt atctttcttc ttccgaaagt gctacttcag aaagagcaag
      601 aactgtacgg ataaggataa gtatatcttc tatctactac aatacgaaaa gaactctcga
      661 actctccctc ccactttact ttaactaata gattatgcag attttcgtca agactttgac
      721 cggtaaaacc ataacattgg aagttgaatc ttccgatacc atcgacaacg ttaagtcgaa
      781 aattcaagac aaggaaggta tccctccaga tcaacaaaga ttgatctttg ccggtaagca
      841 gctagaagac ggtagaacgc tgtctgatta caacattcag aaggagtcca ccttacatct
      901 tgtgctaagg ctaagaggtg gttatcacgg atcctctaga gtcgacctgc aggcatgcaa
      961 gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1021 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1081 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     1141 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     1201 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1261 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1321 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1381 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     1441 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1501 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     1561 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1621 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     1681 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     1741 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1801 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     1861 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1921 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1981 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2041 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2101 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2161 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2221 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2281 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2341 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2401 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2461 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2521 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2581 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2641 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2701 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2761 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2821 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2881 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3061 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3181 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3661 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3721 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3781 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
