LOCUS       sequence-421002-        6567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Gatad2a-exon into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6567)
  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 6567)
  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 Gatad2a-exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(461..508)
                     /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     640..659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    792..1016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1005..1022
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1064..1088
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2904..2928)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2929..2952)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3104..3128)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3150..3168
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3187..3205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3224..3242
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3261..3279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3478..3497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3572..3676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3677..3745,3746..4537)
                     /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(3895..3914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4708..5296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5197..5216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5450..5467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5584..5605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5620..5625,5626..5643,5644..5650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5658..5674
                     /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     5663..5685
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5682..5698
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5682..5698
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5747..5868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5758..5777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5812..5831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5998..6567,1..453))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
       61 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg cgaccggctg
      121 cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg
      181 ggagatgcaa taggtcaggc tctcgctgaa ctccccaatg tcaagcactt ccggaatcgg
      241 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
      301 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
      361 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
      421 ctcgacagac gtcgcggtga gttcaggctt tttggccaag gaagttccta tactttctag
      481 agaataggaa cttcggaata ggaacttcta ggtacgtgaa ccatcaccct aatcaagttt
      541 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
      601 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
      661 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
      721 gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc tggttctttc
      781 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
      841 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
      901 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
      961 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     1021 ggctgatcag cgggtttaaa cgggccctct agactcgagt cagaccactt tgtacaagaa
     1081 agttgggtac tatttccatg tggctgattg ggggatggag cgtggaggaa tcatgtccag
     1141 gaggtattct cgctgtcgat ccacagccga ggaggtcttg tgaactgcca agcttgggct
     1201 gacgaaggcg agggtcccac ctgtgctcag aggcgtcttt ttccagttgg aggcggtgcc
     1261 cttgccggtg cccacgactc tctcggaatg tctgccagtc ctgcttacca gggccgtagc
     1321 tgaggctgca ttctgcagtt tgggtgactg gctgaaagtg tgcagaacac cacgaggtgc
     1381 tgtggccgag cctcgggaca gctggctgga agcctgcttc agcgtgggat gtggagctgc
     1441 aggctcagcc ttgatgctgg cggtacccac accctgctgc agcaggcgct gctccatctc
     1501 ctgctcttgc tgtagagcct tcacaaaggc ggccttcagg cgactggtgt gctccacctt
     1561 gagtgccttc ttttggttgg atgtcatgca gttctcacac ataacggctc cacccttctc
     1621 tcgccagcgg catgtgaagt ctgtcttaca ctgcacacac atgtagggct cccgggataa
     1681 cacagcagct gctgcagtgg cagaaatcct gcctgcttgt gtctccagca ggttctgcac
     1741 cacctcctcc aggccgacca ggtagataaa ctcattattg gcagcactgg gcaagaagtt
     1801 catctctggg gcagggggct tgggtggggg gatctccagc agtgtcttct ccagctgctt
     1861 gcgcagtgct agcttggcag ccgcctgtcg gctagctgga gactcagcag atgcaaccac
     1921 agaagccaca ctggtgggag tggaatttgc ctgagcagaa gcaactgctg tccccttcat
     1981 ggaggctgca gtaggctgag ggatgttgac aagcagactg gtgttgggga cattggcgac
     2041 acggataagt ccctgctgga tgatcctctg tccctgaatc tgcatgctgg gcactgaggc
     2101 acggggggcc aggaggaggg gtggtggccc cgtagaggaa tgctgtctga tgttgtgaat
     2161 ctgggccccc ctgacaagcg gtggcatgac tacttgagag ctggcctgtg gtcccagttt
     2221 ggcagacacc tgctgcccac ctcggaccag tggtggtggg atgatgctgc cgggtattcg
     2281 agtggaggag gtctgaagtg aggtcttgcc agcaggaatg ttctgggtac cccgcacgag
     2341 tgggggaggg gtggtcacag tgctccctga ggaagctgta ggcttctgtg cggtggcttc
     2401 cttctgtatc tgactctgcc gcagcttctt caacaagacc agttttgctt cctctagcct
     2461 cagctcctcc ttcagctgct tgatcatccg ctctcgttct tctgggctgc ttttcaagag
     2521 cgcctcagtg ctggtgtctt tcaaggctac tgtggtcagg ccattcaccc ttgggctcga
     2581 tggctgctca ctgtcagaga gcacaatcac atcaggcgag ggaggtctct tctcagactt
     2641 catgtcactg tgtgaagtgc gcatgtccac aggcccgtct cctaacagcc cctcgccaga
     2701 tcctgtgccc atggcctctg ttgtcctcag caacccctgg gctgagcctg cactcggctc
     2761 tggcatcacc ctgctgtctc cgtccactgt cagctctgat gaccctttct ccattttcat
     2821 tttcttgttc tccacatcat cctccgttag gtcaggttca agggtgcgct tctgactgcg
     2881 ggttcggcat gcttcttcac tggtgccaac ttttttgtac aaacttgtct tgtcatcgtc
     2941 atccttgtaa tcgatgtcat gatcatatgt atcaccgtca tggtctttgt agtccatggt
     3001 ggaagcttaa gtttaaacgc tagaggatcc agcctccgcg gaggctggat cggtcccggt
     3061 gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta
     3121 aacgagctct gcttatatag ggggtacctt ctctatcact gatagggagc tcgagacgtg
     3181 tcgacttctc tatcactgat agggagacgt tctagacgta ccttctctat cactgatagg
     3241 gagctcgaga cgtgtcgact tctctatcac tgatagggag acgtgcggcc agcttggatc
     3301 cctcgagacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa
     3361 gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg
     3421 cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg
     3481 catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc
     3541 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3901 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5041 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5101 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5161 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5221 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5281 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5341 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5401 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5461 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5521 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     5581 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     5641 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     5701 tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag atacattgat
     5761 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     5821 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa
     5881 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     5941 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgta gtacgtgcta
     6001 ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga gtacttctac
     6061 acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc
     6121 ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat catcgaaatt
     6181 gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat acgcccggag
     6241 ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct gctgctccat
     6301 acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg aatccccgaa
     6361 catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt
     6421 ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat
     6481 cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca cagtttgcca
     6541 gtgatacaca tggggatcag caatcgc
//
