LOCUS       sequence-462329-        8075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express S.cerevisiae spt5 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8075)
  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 8075)
  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: pZH60 pLIB-spt5
FEATURES             Location/Qualifiers
     source          1..8075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4048..4081)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4129..4220
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4145..4162
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    7584..7718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7621..7640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7675..7694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7828..7993
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctattcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttt
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cagctttgtt tgtactatca acaggttgaa ctgctgatca
     4021 acagatcctc tacgcggccg cggtaccata acttcgtata gcatacatta tacgaagtta
     4081 tctggtttta ctagtatcga ttcgcgacct actccggaat attaatagat catggagata
     4141 attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg
     4201 taataaaaaa acctataaat attccggatt attcataccg tcccaccatc gggcgcggat
     4261 ccatgagtga caactcggac acaaacgtga gcatgcagga ccatgatcaa caatttgctg
     4321 atcccgtagt ggttccgcag tcaactgaca ctaaagatga aaatactagt gacaaagata
     4381 ctgttgatag tggcaatgtg accacaacgg aaagtacaga acgtgcagaa agtacaagca
     4441 atattccccc tttagatggg gaagaaaaag aagcaaaatc tgagccacag caacctgagg
     4501 ataatgcaga aacggcggcc acagagcagg tttcatctag taacgggcct gctacagatg
     4561 atgcccaagc aactttgaat acggattcat ccgaagcaaa tgaaattgtc aagaaggaag
     4621 agggctctga tgaaaggaag agacctcgcg aagaggacac caaaaacagt gatggtgata
     4681 ccaaagatga gggcgataac aaagatgaag acgatgatga agacgacgat gacgatgatg
     4741 atgatgagga cgatgacgat gaagccccaa ctaaaaggcg tcgtcaggag aggaacagat
     4801 tcttggatat tgaagctgag gttagtgatg atgaagatga agatgaagat gaagaggatt
     4861 cagagttggt tcgtgaaggt ttcattaccc atggtgatga tgaagatgac gaagcaagtg
     4921 ctccaggcgc aagaagagac gatagattac atagacaact ggaccaagat ttgaacaaga
     4981 cttcagaaga agacgctcaa aggttagcga aagaattaag ggagcgttac ggtagaagca
     5041 gctccaagca ataccgtgct gctgctcaag atggttacgt gccccagagg tttctcctac
     5101 caagtgttga tacagctacc atttggggtg tgcgctgcag accaggtaaa gaaaaggaat
     5161 tgattcgtaa gttattaaaa aaaaaattca atttggatag ggcgatgggt aagaaaaaac
     5221 tgaaaatttt atccattttc caaagggata attacacagg aagaatctat atcgaagccc
     5281 ctaagcaatc cgttattgaa aaattttgta atggtgttcc agatatttat atttctcaaa
     5341 aattgctaat tcctgtccaa gaattacctc tattactaaa accaaacaaa tctgatgatg
     5401 ttgctttgga agaaggtagc tacgttcgta ttaagagagg gatctataag ggtgacttag
     5461 ctatggtcga ccaaattagt gagaataatt tagaagttat gctgaaaatt gttcctcgtc
     5521 tggattatgg taaattcgac gaaattgatc caacaacaca gcaacgtaaa tccagaagac
     5581 caacttttgc tcatagagca ccaccgcaat tatttaatcc aacaatggct ctaagattag
     5641 accaagctaa cctgtacaaa agggatgatc gccactttac ttataagaat gaagattata
     5701 tcgatggtta tctgtataag tccttcagaa ttcaacatgt ggaaaccaaa aatattcagc
     5761 caactgtgga ggaattggca agattcggtt ccaaagaggg cgcggtagat ctaacatcag
     5821 tttcacaatc aatcaagaag gctcaagctg caaaggtcac tttccagcca ggtgatcgta
     5881 tcgaagttct aaatggtgaa caacgtggtt ccaagggtat tgtaacaaga accacgaaag
     5941 atattgctac tatcaaacta aatggcttta caacgcctct agaatttcct atctctactc
     6001 tgaggaaaat tttcgaacct ggtgatcacg ttactgtcat caatggtgaa catcaaggtg
     6061 atgctggttt agttcttatg gtagagcaag gtcaagtgac atttatgtca actcaaacaa
     6121 gcagagaagt taccattaca gcaaataatc tgtccaaatc catcgacact acagctacat
     6181 caagtgaata tgcgctacat gacatagtcg aattgagtgc taaaaatgtt gcttgtatta
     6241 ttcaagctgg ccacgatatc ttcaaggtta ttgatgaaac tggtaaagtg tcgacaatta
     6301 cgaaggggtc tatcttgagt aaaataaata ctgctcgtgc acgcgtttct agtgtcgatg
     6361 caaatggtaa tgaaatcaaa attggtgata ccatagtaga gaaagtaggt tcacgcagag
     6421 aaggtcaggt cctgtacata caaacacaac aaatttttgt tgtttcaaag aagattgttg
     6481 aaaatgcagg ggtttttgtt gttaacccta gtaacgttga ggccgtggca tccaaggaca
     6541 atatgttaag taacaaaatg gatctaagta aaatgaatcc agaaatcatt tctaaaatgg
     6601 gacctccatc atctaagaca ttccaacaac ccatccagtc cagaggaggt cgtgaagttg
     6661 cactcggcaa aacagtaaga attcgttctg ctggttacaa gggtcaatta ggtattgtga
     6721 aagatgtgaa tggtgataaa gctactgtcg aattacactc gaagaacaaa cacattacaa
     6781 ttgacaagca taagttaaca tattacaacc gtgagggagg tgaaggtatc acgtatgatg
     6841 aattggttaa tagacgtggt agagttccac aggccagaat gggcccaagt tacgtcagtg
     6901 ccccaagaaa catggccact ggcggtattg cagcaggtgc tgcggctacc tcttctggtc
     6961 ttagcggtgg tatgacacca ggatggagct ccttcgatgg tggcaaaaca ccagctgtaa
     7021 atgcgcatgg aggctcaggt ggtggcggtg tctcctcatg gggtggtgct tccacttggg
     7081 gtggccaagg taatggaggt gcatccgctt ggggcggtgc tggcggcggt gcctcagctt
     7141 ggggcggcca aggtactggt gctacttcta cttggggtgg tgcttcagcc tggggtaaca
     7201 aatcaagttg gggcggtgca tccacttggg cgtcgggtgg tgaatctaat ggtgccatgt
     7261 ctacttgggg tggtaccggt gataggtcag cctacggcgg ggcttccacc tggggaggaa
     7321 ataacaataa taaaagtaca agagatggcg gagcttctgc atggggtaac caagacgatg
     7381 gaaataggtc tgcttggaac aaccaaggaa ataagtcaaa ctatggtggt aacagtacat
     7441 ggggaggtca ttaaaagctt gtcgagaagt actagaggat cataatcagc cataccacat
     7501 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     7561 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     7621 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7681 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagaa
     7741 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     7801 taactatcat aacccctagg agatccgaac cagataagtg aaatctagtt ccaaactatt
     7861 ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca tctattttgt
     7921 cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt
     7981 ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca
     8041 actccatgtg acaaaccgtc atcttcggct acttt
//
