LOCUS       sequence-462092-        3208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid containing dual G-less cassette for in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3208)
  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 3208)
  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: pZH1 Dual G-less cassette
FEATURES             Location/Qualifiers
     source          1..3208
                     /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"
     primer_bind     complement(987..1003)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(987..1003)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1000..1022)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1011..1027
                     /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(1035..1041,1042..1059,1060..1065))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1080..1101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1218..1235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1389..1977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1469..1488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2148..2939,2940..3008))
                     /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     2771..2790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3009..3113)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3181..3199
                     /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 cggatcccgg tgcggccgct ccgatcgatg gcctcgagta atctactcat ctactctact
      841 tcatctatat ttcacacatc tataatcatt cactctcatc ctctctacct tcactcacat
      901 tctattctac tcctctccct ctccatatcc actccaccac tcttcctctc ctctatcaat
      961 cttatcacaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1021 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1081 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1141 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1201 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3121 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3181 aataggcgta tcacgaggcc ctttcgtc
//
