LOCUS       sequence-461128-        7313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7313)
  AUTHORS   Lopez Martinez D, Todorovski I, Noe Gonzalez M, Rusimbi C, Blears D,
            Khallou N, Han Z, Dirac-Svejstrup AB, Svejstrup JQ
  TITLE     PAF1C-mediated activation of CDK12/13 kinase activity is critical 
            for CTD phosphorylation and transcript elongation.
  JOURNAL   Mol Cell. 2025 May 15;85(10):1952-1967.e8. doi: 
            10.1016/j.molcel.2025.04.012. Epub 2025 May 1.
  PUBMED    40315851
REFERENCE   2  (bases 1 to 7313)
  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.2025.04.012"; journalName: "Mol Cell"; date: 
            "2025-05-15- 15"; volume: "85"; issue: "10"; pages: "1952-1967.e8"
COMMENT     Sequence Label: pLIB-FLAG(N)-CDK12deltaNTD(700-1490)
FEATURES             Location/Qualifiers
     source          1..7313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1478..1612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1515..1534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1569..1588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1722..1887
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2071..2525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2158..2177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2367..2388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2551..2655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2656..2724,2725..3516)
                     /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(2874..2893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3687..4275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4176..4195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4429..4446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4580..4804
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4871..5404)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5593..5598,5599..5615,5616..5621))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(6017..6050)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6098..6189
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6114..6131
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             6235..6258
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6265..6288
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 gtggagtaag aaacggcgac gtcagcgaca aagtggtgtt gtagtcgaag agccacctcc
       61 atccaaaact tctcgaaaag aaactacctc agggacaagt actgagcctg tgaagaacag
      121 cagcccagca ccacctcagc ctgctcctgg caaggtggag tctggggctg gggatgcaat
      181 aggccttgct gacatcacac aacagctgaa tcaaagtgaa ttggcagtgt tattaaacct
      241 gctgcagagc caaaccgacc tgagcatccc tcaaatggca cagctgctta acatccactc
      301 caacccagag atgcagcagc agctggaagc cctgaaccaa tccatcagtg ccctgacgga
      361 agctacttcc cagcagcagg actcagagac catggcccca gaggagtctt tgaaggaagc
      421 accctctgcc ccagtgatcc tgccttcagc agaacagacg acccttgaag cttcaagcac
      481 accagctgac atgcagaata tattggcagt tctcttgagt cagctgatga aaacccaaga
      541 gccagcaggc agtctggagg aaaacaacag tgacaagaac agtgggccac aggggccccg
      601 aagaactccc acaatgccac aggaggaggc agcagcatgt cctcctcaca ttcttccacc
      661 agagaagagg ccccctgagc cccccggacc tccaccgccg ccacctccac cccctctggt
      721 tgaaggcgat ctttccagcg ccccccagga gttgaaccca gccgtgacag ccgccttgct
      781 gcaactttta tcccagcctg aagcagagcc tcctggccac ctgccacatg agcaccaggc
      841 cttgagacca atggagtact ccacccgacc ccgtccaaac aggacttatg gaaacactga
      901 tgggcctgaa acagggttca gtgccattga cactgatgaa cgaaactctg gtccagcctt
      961 gacagaatcc ttggtccaga ccctggtgaa gaacaggacc ttctcaggct ctctgagcca
     1021 ccttggggag tccagcagtt accagggcac agggtcagtg cagtttccag gggaccagga
     1081 cctccgtttt gccagggtcc ccttagcgtt acacccggtg gtcgggcaac cattcctgaa
     1141 ggctgaggga agcagcaatt ctgtggtaca tgcagagacc aaattgcaaa actatgggga
     1201 gctggggcca ggaaccactg gggccagcag ctcaggagca ggccttcact gggggggccc
     1261 aactcagtct tctgcttatg gaaaactcta tcgggggcct acaagagtcc caccaagagg
     1321 gggaagaggg agaggagttc cttactaaaa gcttgtcgag aagtactaga ggatcataat
     1381 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1441 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     1621 gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     1681 ccaccgctga gcaataacta tcataacccc taggagatcc gaaccagata agtgaaatct
     1741 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     1801 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     1861 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat
     1921 caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac
     1981 agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     2041 cgcttatttg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     2101 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2161 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2221 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2281 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2341 tgacgttgga gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa
     2401 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2461 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2521 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2641 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2701 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2761 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2821 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2881 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2941 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3001 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3061 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3121 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3181 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3241 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3301 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3361 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3421 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3481 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tattcttcta
     3841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3961 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4141 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4201 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4261 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4321 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4381 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4441 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4501 tcacaccgca tagaccagcc gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg
     4561 gatgccctgc gtaagcgggt gtgggcggac aataaagtct taaactgaac aaaatagatc
     4621 taaactatga caataaagtc ttaaactaga cagaatagtt gtaaactgaa atcagtccag
     4681 ttatgctgtg aaaaagcata ctggactttt gttatggcta aagcaaactc ttcattttct
     4741 gaagtgcaaa ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg taaagactat
     4801 attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt
     4861 gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta
     4921 tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga
     4981 tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa
     5041 tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg
     5101 gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga
     5161 aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg
     5221 gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag
     5281 cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg
     5341 agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta
     5401 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
     5461 gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
     5521 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     5581 tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggtttg tgccttcatc
     5641 cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg
     5701 tcctggctgg cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc
     5761 ttgctgttct tctacggcaa ggtgctgtgc acggatctgc ccttgcttca ggagatcggt
     5821 agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc
     5881 ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt
     5941 ggttggctac agctttgttt gtactatcaa caggttgaac tgctgatcaa cagatcctct
     6001 acgcggccgc ggtaccataa cttcgtatag catacattat acgaagttat ctggttttac
     6061 tagtatcgat tcgcgaccta ctccggaata ttaatagatc atggagataa ttaaaatgat
     6121 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
     6181 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc catggactac
     6241 aaggacgacg atgacaagct cgatcttgaa gtcctctttc agggacccat gccaaaaatt
     6301 tgttgtcctc gttatggaga aagaagacaa acagaaagcg actgggggaa acgctgtgtg
     6361 gacaagtttg acattattgg gattattgga gaaggaacct atggccaagt atataaagcc
     6421 aaggacaaag acacaggaga actagtggct ctgaagaagg tgagactaga caatgagaaa
     6481 gagggcttcc caatcacagc cattcgtgaa atcaaaatcc ttcgtcagtt aatccaccga
     6541 agtgttgtta acatgaagga aattgtcaca gataaacaag atgcactgga tttcaagaag
     6601 gacaaaggtg ccttttacct tgtatttgag tatatggacc atgacttaat gggactgcta
     6661 gaatctggtt tggtgcactt ttctgaggac catatcaagt cgttcatgaa acagctaatg
     6721 gaaggattgg aatactgtca caaaaagaat ttcctgcatc gggatattaa gtgttctaac
     6781 attttgctga ataacagtgg gcaaatcaaa ctagcagatt ttggacttgc tcggctctat
     6841 aactctgaag agagtcgccc ttacacaaac aaagtcatta ctttgtggta ccgacctcca
     6901 gaactactgc taggagagga acgttacaca ccagccatag atgtttggag ctgtggatgt
     6961 attcttgggg aactattcac aaagaagcct atttttcaag ccaatctgga actggctcag
     7021 ctagaactga tcagccgact ttgtggtagc ccttgtccag ctgtgtggcc tgatgttatc
     7081 aaactgccct acttcaacac catgaaaccg aagaagcaat atcgaaggcg tctacgagaa
     7141 gaattctctt tcattccttc tgcagcactt gatttattgg accacatgct gacactagat
     7201 cctagtaagc ggtgcacagc tgaacagacc ctacagagcg acttccttaa agatgtcgaa
     7261 ctcagcaaaa tggctcctcc agacctcccc cactggcagg attgccatga gtt
//
