LOCUS       sequence-421246-        8491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Camta2-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8491)
  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 8491)
  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 Camta2-exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..8491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..148
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    190..214
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3806..3830)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3831..3854)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4006..4030)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4052..4070
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4089..4107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4126..4144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4163..4181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4200..4218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4237..4255
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4274..4292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4311..4329
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4528..4547
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4622..4726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4727..4795,4796..5587)
                     /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(4945..4964)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5758..6346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6247..6266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6500..6517
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6634..6655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6670..6675,6676..6693,6694..6700)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6708..6724
                     /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)."
     primer_bind     6713..6735
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6732..6748
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6732..6748
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6797..6918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6808..6827)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6862..6881
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7048..8070)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(8078..8125)
                     /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)."
     primer_bind     8257..8276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    join(8409..8491,1..142)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
       61 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
      121 ggctggcaac tagaaggcac agtcgaggct gatcagcggg tttaaacggg ccctctagac
      181 tcgagtcaga ccactttgta caagaaagtt gggtatcagg tggccagtcc tggctggggc
      241 agcccctcca gctcctggtt ctgcttcagt tccctcattc tgtgtcggca gcgccgcagg
      301 aacctcatta ttttccgggc tgcctggtct tgcttcttgg tgagaaaggt gcccttgttt
      361 cgagcaggaa gggggcctga gggccggtga ggaggccccg gcctgcggcg gtaggagcgg
      421 tagtgctgct ggataagcac cgctgctcgg cggctctgct gaaaccgctt ctgttcatag
      481 tagcttcgga acttgctctg aatcaggatg gccgcctgag tcattttctt atagagtgca
      541 aactgcttgt acttccgata acaacgttgg atcaccgctg cggctacctc ctgctgctcc
      601 ttcagcctcc gacccttgta ctttcggaaa gctgtctgga tgactcgtgc tgcctcatac
      661 agctcccgct gctcgtgatc tgagagagtc agcagggcaa aatcactttc catcttgcca
      721 ctggtagatg cagagaggaa ctctgcccag gaaggtgcag gggggatagc gaggcgacct
      781 cgttcaaagg gcagctcact gggaggagct gagctgggga aatgatccac attctccagg
      841 taactggcca gccaggacat ggtctcactg agccccacgg tgcctgtgtg ctcccgtggt
      901 gaggctccag cgtcagggaa ctctgagaag tcctctcgtt taatccgctc tggtgtggcg
      961 tcgatgatct gcttggccag tgagatcatg tccaccggga tcacatccac agctggtggg
     1021 ctgtcagcat cctctggagc agccccattg tcctgggcta gtggggggcc gcacggggag
     1081 ggagcaggtt ctttggagtt agtggcttca tagtccatga ggagtaaggg tgtctcaggg
     1141 gcaccacaag aaagctggcc tgggatcatc tccatagaaa tgtcagaggc cagcggagga
     1201 gcagggggag gactgccatc tggggcactt gagtaggctg atgtcacaga gaaagtgcca
     1261 tctgacagct cggagggtga cgaggcgctg ctcaggccag tgtctgggct agaggatggt
     1321 ggggatagag cgagtgggtg ctcaactgaa agctcctgtc tctgcagttc ctcaaggcag
     1381 cgggcaaggc gcacgtgacc acgagaatga gccacagaca gggggagtcg gcccagagag
     1441 tccggaatgc tcagtgcttg tcggttccaa cagaaaagaa gcacagcagc ttctaagtgt
     1501 cccagagcac aggcccacat cagaggggtg caagagaaat ggtccacatt gagcgggtca
     1561 acctcttgct ctaagtccaa gcttcctgtt tccacactcc tccactggct cagagtctcg
     1621 atgagccgag cgtagccctg tgcagcagcc agatgtagga ggctcatgcc ccggaaggga
     1681 cttccatgga tcagccgttc aggacccctc caagtggacc gcgggatcat gctctctacc
     1741 aagaccacca cccgtgcctc gaagccaggg ccctggcctt catcctgaat tggaggagcc
     1801 tctgggccct gaccaggagc ctgcccagct gccgcaatct ctgccatccg cttctccatc
     1861 tgctccagcc gctccaggat ggacatccgg aactggctgt catccagtga caaccagtcg
     1921 agctgtgtac taggcagaga caggaaccgg cgggctcgat actcaaagag cacagaggca
     1981 gagagggggc cctctcgccc cgccacctgc aaagacacca gccctacctc atgggcagga
     2041 cagtagcagc gtaagacacc aggctggacc agggaggctg gcactgcgat gtgatcgaag
     2101 acacaggagt aatgctctgc ggcctctgtc caagggcctg tgatgagcac cttgaccccg
     2161 ccctctgggt aggaccactc tggggagaag tctgtgatgg catttagcgc aggagagagc
     2221 tggggggcgg gggcagggac acctggagct tcatcactga tgagttctcc catgaggtct
     2281 gggaatgacg aaagactgaa gggctcgagc tcgctgctgc tgccagcaga gcctccgaac
     2341 aaggcttctc ctctcgttgc cctgcctgat ggcggcaagg cggttgggga ggacggggat
     2401 gaaggagggg atgaaggtac cggcggcacc gttcccggac ccttgagttc ctccccacta
     2461 tcatcatctt ggataaagaa tttgtttcct cttctcccgt ttgctcccaa ctgtggagcg
     2521 agacctcgag cagccgcctg gggctccagg gcagctgcag gttccagggc aggacaggga
     2581 gtgtgagtgc cctctgcctc ggggaagtct gggtttactc cttggccccc tccgtatgtc
     2641 tggccccttt gggggctgtt gagaaaacga tcagggtcaa aggcagggct gggaggagcc
     2701 ggtggggcag aaggctcaga acctacaacc acggccaagc tcacaggagg tctaggctca
     2761 gcctgcggag ccaagtgcct ggtgggtgtc aggcccccag cgcgctgctc tagtcctgtt
     2821 aggaggagga tagcagtgcc tcctcttgat gaaccccctc gagaggtagg agggctaggc
     2881 ctgatttcta ggggttctgc aaatcctgag gaggaggagg aagaagagga ggaagatggg
     2941 gaggtgtgtg ccttggggag ctctggagga agtggagcta tcagtggtgg gggttcgggg
     3001 ggcttggagt gggatataga agccagggct aaagctcggg gctccacttt gggagagatg
     3061 atgcggtgtt tcgtgctact gcatttgtgg gtaaggctcc cggaaccaag gcccccactg
     3121 cagagacaag catgagttct gggtgctggc ttggttgggt gggtgtccaa gatctgttgc
     3181 accaactgtt ccacggagaa ttcctctgcc ccattcccac agctccactt gatgccatga
     3241 aacatgggct tcagctgtcc caagagctcc tcacgtgacc acttcagcca ttctcgacgg
     3301 tcgctactga tggaacaaaa gatggggcta cagccctttc cacaatcctc cagggctggg
     3361 acgttcaggt agtgcacaag gacgatgtcg gggttctgga gcagccaata gcagcgccgg
     3421 tggaatgtgg ggacgatgga agagtgaacg tagcagccat agagacactc catgccctgg
     3481 accttgagtt tcatgtggtc ctctcgggta gtcttcccat ccttccgttt cttccaaagg
     3541 taaccatcct tccggtattt caccttctta cgattgtaga ggataatgga gccattctga
     3601 ggccttgtct ttggggcaca agacagccac tcatcgtgtt tctcaaaggt gatcaggtag
     3661 gatgcaatct cctcatttgt attccatcgg agccgctctg gaggcaacag tgggcatcga
     3721 ggaagacact ccagcaattt cttggggagg aagatcttca ggtggtggct attctcagca
     3781 acctcagtgg tgtccttggt attggtgcca acttttttgt acaaacttgt cttgtcatcg
     3841 tcatccttgt aatcgatgtc atgatcatat gtatcaccgt catggtcttt gtagtccatg
     3901 gtggaagctt aagtttaaac gctagaggat ccagcctccg cggaggctgg atcggtcccg
     3961 gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac
     4021 taaacgagct ctgcttatat agggggtacc ttctctatca ctgataggga gctcgagacg
     4081 tgtcgacttc tctatcactg atagggagac gttctagacg taccttctct atcactgata
     4141 gggagctcga gacgtgtcga cttctctatc actgataggg agacgttcta gacgtacctt
     4201 ctctatcact gatagggagc tcgagacgtg tcgacttctc tatcactgat agggagacgt
     4261 tctagacgta ccttctctat cactgatagg gagctcgaga cgtgtcgact tctctatcac
     4321 tgatagggag acgtgcggcc agcttggatc cctcgagacg cgtatatctg gcccgtacat
     4381 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     4441 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     4501 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4561 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     4621 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4681 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4741 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4801 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4861 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4921 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4981 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5041 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5101 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5161 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5221 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5281 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5341 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5401 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5461 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5521 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5581 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5641 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5701 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5761 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5821 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5881 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5941 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6001 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6061 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6121 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6181 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6241 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6301 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6361 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6421 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6481 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6541 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6601 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6661 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6721 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     6781 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     6841 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     6901 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     6961 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     7021 ggtggaatcg aaatctcgta gtacgtgcta ttcctttgcc ctcggacgag tgctggggcg
     7081 tcggtttcca ctatcggcga gtacttctac acagccatcg gtccagacgg ccgcgcttct
     7141 gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg
     7201 accctgcgcc caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc
     7261 aagaccaatg cggagcatat acgcccggag ccgcggcgat cctgcaagct ccggatgcct
     7321 ccgctcgaag tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat
     7381 gttggcgacc tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt
     7441 tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg
     7501 gacttcgggg cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc
     7561 actgacggtg tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca
     7621 tatgaaatca cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc
     7681 gctcgtctgg ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga ccggctgcag
     7741 aacagcgggc agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga
     7801 gatgcaatag gtcaggctct cgctgaactc cccaatgtca agcacttccg gaatcgggag
     7861 cgcggccgat gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct
     7921 atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc
     7981 gccctccgag agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc
     8041 gacagacgtc gcggtgagtt caggcttttt ggccaaggaa gttcctatac tttctagaga
     8101 ataggaactt cggaatagga acttctaggt acgtgaacca tcaccctaat caagtttttt
     8161 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     8221 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     8281 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     8341 taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg ttctttccgc
     8401 ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     8461 ccccttgctg tcctgcccca ccccaccccc c
//
