LOCUS       sequence-420962-        6873 bp    DNA     circular SYN 15-APR-2025
DEFINITION  LR vector for integration of Tcf12+exon(72nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6873)
  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 6873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..6873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    713..834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(750..769)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     804..823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(883..899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(883..899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(896..918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    907..923
                     /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(931..937,938..955,956..961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    976..997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1114..1131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1285..1873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1365..1384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2044..2835,2836..2904))
                     /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     2667..2686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2905..3009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3084..3103)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3302..3320
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3339..3357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3376..3394
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3413..3431
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3453..3477
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3629..3652
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3653..3677
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5799..5823)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5865..5882)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5871..6095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6228..6247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6379..6426
                     /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(6434..6873,1..583)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
       61 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
      121 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
      181 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
      241 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
      301 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
      361 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
      421 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
      481 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
      541 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt
      601 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
      661 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
      721 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
      781 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
      841 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
      901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1021 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1081 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1201 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1261 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1321 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1381 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1441 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1501 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1621 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1681 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1741 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1801 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     3061 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3121 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     3181 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     3241 ctgcttcgcg atgtacgggc cagatatacg cgtctcgagg gatccaagct ggccgcacgt
     3301 ctccctatca gtgatagaga agtcgacacg tctcgagctc cctatcagtg atagagaagg
     3361 tacgtctaga acgtctccct atcagtgata gagaagtcga cacgtctcga gctccctatc
     3421 agtgatagag aaggtacccc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     3481 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
     3541 cgcggaggct ggatcctcta gcgtttaaac ttaagcttcc accatggact acaaagacca
     3601 tgacggtgat acatatgatc atgacatcga ttacaaggat gacgatgaca agacaagttt
     3661 gtacaaaaaa gttggcacca atccccagca gcagcgcatg gccgcgatag ggaccgacaa
     3721 ggagctgagc gacctgctgg acttcagtgc gatgttttct ccacctgtta atagtgggaa
     3781 aacgagacca acaacactgg gaagtagtca gttcagcggg tcaggtatgg atgagcgtgg
     3841 aggaacaaca tcttggggaa caagtggtca accaagcccc tcctatgatt catctagagg
     3901 ttttacagac agccctcatt acagtgatca cttgaatgac agtcgattag gaacccacga
     3961 aggcttgtcc ccaacacctt tcatgaactc aaatctgata gggaaaacat cagagagagg
     4021 ctcattttcc ctgtacagca gagactctgg actctcaggc tgtcagtcta gtctcctgag
     4081 acaagatcta ggacttggga gcccggcaca gctgtcttct tcaggaaagc ctggaacacc
     4141 atactactca ttctctgcca caagttcgag aagaagaccg ctccatgatt ctgtagccct
     4201 agatccttta caagcgaaga aagtaagaaa ggtgcctcct ggcctacctt cttctgtata
     4261 tgcaccatcc ccaaattctg acgatttcaa ccgtgaatct cctagttacc catctcccaa
     4321 gccaccaacc agtatgttcg ctagcacttt ctttatgcaa gatgggaccc acagttcttc
     4381 tgacctttgg agttcatcga atgggatgag ccagcctggt tttggtggaa ttctggggac
     4441 ctccacatcc cacatgtctc agtccagtag ttatggcagc cttcattcac atgaccgctt
     4501 gagttatcct ccacactcag tgtcaccaac agacataaac acaagtcttc cgccaatgtc
     4561 cagcttccac cgtggtagta ccagcagctc accatatgtt gccgcctcac atactcctcc
     4621 catcaatgga tcagatagca tcctaggaac cagagggaat gctgctggaa gctcacagac
     4681 gggtgatgca cttgggaagg ccttggcatc tatttattcc cctgaccaca caagcagtag
     4741 ttttccatca aacccatcaa caccagtggg atctccttca cctctcacag gtaccagtca
     4801 gtggcccaga gctggagggc aagctccttc atctccaagc tatgaaaact cacttcactc
     4861 cctgaaaaat cgagttgagc agcaacttca cgagcatttg caagatgcaa tgtccttctt
     4921 aaaggatgtc tgtgagcagt ctcgaatgga agaccgctta gacaggctgg atgatgctat
     4981 ccatgtgcta cgaaaccatg cagttggacc ttctaccagt ctgcctacta gccacagtga
     5041 catacacagt ttgctgggac catcccataa tgcatcaatt ggaaacctca attcaaacta
     5101 tggaggatcc agccttgtta caaatagtcg atcagcttcg atggtcggaa cacatcggga
     5161 agattcagtc agtctcaatg gcaatcattc ggtcctgtct agtactgttg ctgcctcaaa
     5221 cacagaactg aaccataaaa caccagaaaa tttcagaggt ggtgtacaaa atcagtctgg
     5281 aagtgttgtt ccaacagaaa tcaagactga aaacaaagaa aaagatgaaa accttcatga
     5341 acctccttca tcagatgaca tgaaatcaga tgatgagtcc tcccagaaag acatcaaggt
     5401 ctcatctagg ggcagaacaa gcagtaccaa tgaagacgag gatctgaatc cagaacagaa
     5461 aatcgaaagg gagaaggaaa ggcggatggc taacaatgcc agagagcgcc tgcgcgtgcg
     5521 ggatattaac gaggcgttca aggagcttgg ccgaatgtgt cagcttcatt tgaagagtga
     5581 aaaacctcag acaaaacttc tcattcttca tcaggccgtg gcagtcatcc ttagtctaga
     5641 acagcaagtg agagagagga acctcaaccc caaagcagcc tgccttaaga gaagagaaga
     5701 agaaaaagtc tctgctgcgt cagcagagcc gcccagcacg ttgccaggag cccatcctgg
     5761 gcttagtgag tctaccaacc ctatgggtca tctgtaatac ccaactttct tgtacaaagt
     5821 ggtctgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     5881 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5941 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6001 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     6061 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     6121 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6181 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6241 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6301 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6361 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     6421 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     6481 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     6541 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     6601 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     6661 gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     6721 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     6781 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     6841 accgcaagga atcggtcaat acactacatg gcg
//
