LOCUS       sequence-420968-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Tcf12-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  AUTHORS   Roth JF, Braunschweig U, Wu M, Li JD, Lin ZY, Larsen B, Weatheritt 
            RJ, Gingras AC, Blencowe BJ
  TITLE     Systematic analysis of alternative exon-dependent interactome 
            remodeling reveals multitasking functions of gene regulatory 
            factors.
  JOURNAL   Mol Cell. 2023 Dec 7;83(23):4222-4238.e10. doi: 
            10.1016/j.molcel.2023.10.034.
  PUBMED    38065061
REFERENCE   2  (bases 1 to 6801)
  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.2023.10"; journalName: "Mol Cell"; date: 
            "2023-12-7- 7"; volume: "83"; issue: "23"; pages: "4222-4238.e10"
COMMENT     Sequence Label: 5'-3xFLAG Tcf12-exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    677..798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(714..733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     768..787
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(847..863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(847..863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(860..882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    871..887
                     /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(895..901,902..919,920..925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    940..961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1078..1095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1249..1837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1329..1348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2008..2799,2800..2868))
                     /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     2631..2650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2869..2973)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3048..3067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3266..3284
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3303..3321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3340..3358
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3377..3395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3417..3441
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3593..3616
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3617..3641
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5691..5715)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5757..5774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5763..5987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6120..6139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6271..6318
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             join(6326..6801,1..547)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
       61 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
      121 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
      181 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
      241 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
      301 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
      361 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
      421 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
      481 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
      541 ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
      601 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
      661 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
      721 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
      781 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
      841 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
      901 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
      961 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1021 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1081 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1141 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1201 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1261 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1321 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1381 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1441 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1501 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1561 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1621 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1681 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1741 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1801 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1861 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1921 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1981 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2041 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2101 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2161 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2221 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2281 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2341 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2401 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2461 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2521 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2581 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2641 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2701 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2761 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2821 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2881 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2941 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3001 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     3061 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     3121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     3181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtctc
     3241 gagggatcca agctggccgc acgtctccct atcagtgata gagaagtcga cacgtctcga
     3301 gctccctatc agtgatagag aaggtacgtc tagaacgtct ccctatcagt gatagagaag
     3361 tcgacacgtc tcgagctccc tatcagtgat agagaaggta ccccctatat aagcagagct
     3421 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     3481 agacaccggg accgatccag cctccgcgga ggctggatcc tctagcgttt aaacttaagc
     3541 ttccaccatg gactacaaag accatgacgg tgatacatat gatcatgaca tcgattacaa
     3601 ggatgacgat gacaagacaa gtttgtacaa aaaagttggc accaatcccc agcagcagcg
     3661 catggccgcg atagggaccg acaaggagct gagcgacctg ctggacttca gtgcgatgtt
     3721 ttctccacct gttaatagtg ggaaaacgag accaacaaca ctgggaagta gtcagttcag
     3781 cgggtcaggt atggatgagc gtggaggaac aacatcttgg ggaacaagtg gtcaaccaag
     3841 cccctcctat gattcatcta gaggttttac agacagccct cattacagtg atcacttgaa
     3901 tgacagtcga ttaggaaccc acgaaggctt gtccccaaca cctttcatga actcaaatct
     3961 gatagggaaa acatcagaga gaggctcatt ttccctgtac agcagagact ctggactctc
     4021 aggctgtcag tctagtctcc tgagacaaga tctaggactt gggagcccgg cacagctgtc
     4081 ttcttcagga aagcctggaa caccatacta ctcattctct gccacaagtt cgagaagaag
     4141 accgctccat gattctgtag ccctagatcc tttacaagcg aagaaagtaa gaaaggtgcc
     4201 tcctggccta ccttcttctg tatatgcacc atccccaaat tctgacgatt tcaaccgtga
     4261 atctcctagt tacccatctc ccaagccacc aaccagtatg ttcgctagca ctttctttat
     4321 gcaagatggg acccacagtt cttctgacct ttggagttca tcgaatggga tgagccagcc
     4381 tggttttggt ggaattctgg ggacctccac atcccacatg tctcagtcca gtagttatgg
     4441 cagccttcat tcacatgacc gcttgagtta tcctccacac tcagtgtcac caacagacat
     4501 aaacacaagt cttccgccaa tgtccagctt ccaccgtggt agtaccagca gctcaccata
     4561 tgttgccgcc tcacatactc ctcccatcaa tggatcagat agcatcctag gaaccagagg
     4621 gaatgctgct ggaagctcac agacgggtga tgcacttggg aaggccttgg catctattta
     4681 ttcccctgac cacacaagca gtagttttcc atcaaaccca tcaacaccag tgggatctcc
     4741 ttcacctctc acaggtacca gtcagtggcc cagagctgga gggcaagctc cttcatctcc
     4801 aagctatgaa aactcacttc actccctgca gtctcgaatg gaagaccgct tagacaggct
     4861 ggatgatgct atccatgtgc tacgaaacca tgcagttgga ccttctacca gtctgcctac
     4921 tagccacagt gacatacaca gtttgctggg accatcccat aatgcatcaa ttggaaacct
     4981 caattcaaac tatggaggat ccagccttgt tacaaatagt cgatcagctt cgatggtcgg
     5041 aacacatcgg gaagattcag tcagtctcaa tggcaatcat tcggtcctgt ctagtactgt
     5101 tgctgcctca aacacagaac tgaaccataa aacaccagaa aatttcagag gtggtgtaca
     5161 aaatcagtct ggaagtgttg ttccaacaga aatcaagact gaaaacaaag aaaaagatga
     5221 aaaccttcat gaacctcctt catcagatga catgaaatca gatgatgagt cctcccagaa
     5281 agacatcaag gtctcatcta ggggcagaac aagcagtacc aatgaagacg aggatctgaa
     5341 tccagaacag aaaatcgaaa gggagaagga aaggcggatg gctaacaatg ccagagagcg
     5401 cctgcgcgtg cgggatatta acgaggcgtt caaggagctt ggccgaatgt gtcagcttca
     5461 tttgaagagt gaaaaacctc agacaaaact tctcattctt catcaggccg tggcagtcat
     5521 ccttagtcta gaacagcaag tgagagagag gaacctcaac cccaaagcag cctgccttaa
     5581 gagaagagaa gaagaaaaag tctctgctgc gtcagcagag ccgcccagca cgttgccagg
     5641 agcccatcct gggcttagtg agtctaccaa ccctatgggt catctgtaat acccaacttt
     5701 cttgtacaaa gtggtctgac tcgagtctag agggcccgtt taaacccgct gatcagcctc
     5761 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5821 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5881 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5941 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     6001 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     6061 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     6121 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6181 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6241 acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct
     6301 agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     6361 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     6421 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     6481 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     6541 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     6601 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     6661 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     6721 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     6781 gattgctgat ccccatgtgt a
//
