LOCUS       sequence-462104-        3707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast S.cerevisiae CYC1 polyA termination region inserted between 
            the dual G-less cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3707)
  AUTHORS   Han Z, Moore GA, Mitter R, Lopez Martinez D, Wan L, Dirac Svejstrup 
            AB, Rueda DS, Svejstrup JQ
  TITLE     DNA-directed termination of RNA polymerase II transcription.
  JOURNAL   Mol Cell. 2023 Sep 21;83(18):3253-3267.e7. doi: 
            10.1016/j.molcel.2023.08.007. Epub 2023 Sep 7.
  PUBMED    37683646
REFERENCE   2  (bases 1 to 3707)
  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"; doi: 
            "10.1016/j.molcel.2023.08.007"; journalName: "Mol Cell"; date: 
            "2023-09-21- 21"; volume: "83"; issue: "18"; pages: "3253-3267.e7"
COMMENT     Sequence Label: pZH20 Dual G-less containing yCYC1 polyA
FEATURES             Location/Qualifiers
     source          1..3707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      813..1062
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(829..847)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1486..1502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1486..1502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1499..1521)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1510..1526
                     /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)."
     promoter        complement(join(1534..1540,1541..1558,1559..1564))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1579..1600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1717..1734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1888..2476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1968..1987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2647..3438,3439..3507))
                     /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     3270..3289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3508..3612)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3680..3698
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cctcggcggc ttctaatccg
      421 atcggagggc tgtcacccgc tcggagcact gtactccgct cagaggactg tcctccgctc
      481 ggagcagtgc gccccgactc tagagcatcc ccggtgttcc tgaagggggg ctataaaagg
      541 gggtgggggc gcgttcgtcc tcactctctt cccctccata cccttcctcc atctatacca
      601 ccctactctc ctttcctcat tattcctcct attatcttct cctcttctct ccttcttcta
      661 tatttcccaa atctatcatc attcactctc atcccctctt ccttcactcc cattctattc
      721 tactcctttc cctttccata tcccctccac cccccttcct cccctctttc aatcttatcc
      781 cggatccaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
      841 ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
      901 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
      961 caaatttttc ttttttttct gtacaaacgc gtgtacgcat gtaacattat actgaaaacc
     1021 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcaagcttcg cagtttacac
     1081 tctcatcgtc gctctcatca tcgcttccgt tgttgttttc cttagtagcg tctgcttcca
     1141 gagagtattt atctcttatt acctctaaag gttctgcttg atttctgact ttgttcgcct
     1201 catgtgcata tttttcttgg ttcttttggg acaaaatatg cgtaaaggac ttttgttgtt
     1261 ccctcacatt ccagtttagt tgtcgactga gcggccgctc cgatcgatgg cctcgagtaa
     1321 tctactcatc tactctactt catctatatt tcacacatct ataatcattc actctcatcc
     1381 tctctacctt cactcacatt ctattctact cctctccctc tccatatcca ctccaccact
     1441 cttcctctcc tctatcaatc ttatcacaag cttggcgtaa tcatggtcat agctgtttcc
     1501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1561 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1621 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1681 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1741 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1801 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1861 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1921 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1981 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2041 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2101 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2161 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2221 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2281 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2341 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2401 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2581 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2641 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2821 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     2941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3061 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3301 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3481 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3541 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3601 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3661 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
