LOCUS       sequence-421008-        6642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Gatad2a+exon(75nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6642)
  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 6642)
  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..6642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(423..470)
                     /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     602..621
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    754..978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     967..984
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1026..1050
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2941..2965)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2966..2989)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3141..3165)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     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"
     protein_bind    3298..3316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3515..3534
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3609..3713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3714..3782,3783..4574)
                     /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(3932..3951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4745..5333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5234..5253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5487..5504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5621..5642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5657..5662,5663..5680,5681..5687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5695..5711
                     /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     5700..5722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5719..5735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5719..5735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5784..5905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5795..5814)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5849..5868
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6035..6642,1..415))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatcgcat
       61 ccatggcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc aggtcttgca
      121 acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aactccccaa
      181 tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat
      241 ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat
      301 cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt
      361 cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc tttttggcca
      421 aggaagttcc tatactttct agagaatagg aacttcggaa taggaacttc taggtacgtg
      481 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
      541 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
      601 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
      661 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc
      721 tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat
      781 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
      841 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
      901 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      961 ggcaactaga aggcacagtc gaggctgatc agcgggttta aacgggccct ctagactcga
     1021 gtcagaccac tttgtacaag aaagttgggt actatttcca tgtggctgat tgggggatgg
     1081 agcgtggagg aatcatgtcc aggaggtatt ctcgctgtcg atccacagcc gaggaggtct
     1141 tgtgaactgc caagcttggg ctgacgaagg cgagagtccc acctgtgctc agaggcgtct
     1201 ttttccagtt ggaggcggtg cccttgccgg tgcccacgac tctctcggaa tgtctgccag
     1261 tcctgcttac cagggccgta gctgaggctg cattctgcag tttgggtgac tggctgaaag
     1321 tgtgcagaac accacgaggt gctgtggccg agcctcggga cagctggctg gaagcctgta
     1381 gtgaggaccc gttattccag acacgggcct ctcctgagcg gaacgccaac ttacgtcggg
     1441 gctttatgac ctgcttcagc gtgggatgtg gagctgcagg ctcagccttg atgctggcgg
     1501 tacccacacc ctgctgcagc aggcgctgct ccatctcctg ctcttgctgt agagccttca
     1561 caaaggcggc cttcaggcga ctggtgtgct ccaccttgag tgccttcttt tggttggatg
     1621 tcatgcagtt ctcacacata acggctccac ccttctctcg ccagcggcat gtgaagtctg
     1681 tcttacactg cacacacatg tagggctccc gggataacac agcagctgct gcagtggcag
     1741 aaatcctgcc tgcttgtgtc tccagcaggt tctgcaccac ctcctccagg ccgaccaggt
     1801 agataaactc attattggca gcactgggca agaagttcat ctctggggca gggggcttgg
     1861 gtggggggat ctccagcagt gtcttctcca gctgcttgcg cagtgctagc ttggcagccg
     1921 cctgtcggct agctggagac tcagcagatg caaccacaga agccacactg gtgggagtgg
     1981 aatttgcctg agcagaagca actgctgtcc ccttcatgga ggctgcagta ggctgaggga
     2041 tgttgacaag cagactggtg ttggggacat tggcgacacg gataagtccc tgctggatga
     2101 tcctctgtcc ctgaatctgc atgctgggca ctgaggcacg gggggccagg aggaggggtg
     2161 gtggccccgt agaggaatgc tgtctgatgt tgtgaatctg ggcccccctg acaagcggtg
     2221 gcatgactac ttgagagctg gcctgtggtc ccagtttggc agacacctgc tgcccacctc
     2281 ggaccagtgg tggtgggatg atgctgccgg gtattcgagt ggaggaggtc tgaagtgagg
     2341 tcttgccagc aggaatgttc tgggtacccc gcacgagtgg gggaggggtg gtcacagtgc
     2401 tccctgagga agctgtaggc ttctgtgcgg tggcttcctt ctgtatctga ctctgccgca
     2461 gcttcttcaa caagaccagt tttgcttcct ctagcctcag ctcctccttc agctgcttga
     2521 tcatccgctc tcgttcttct gggctgcttt tcaagagcgc ctcagtgctg gtgtctttca
     2581 aggctactgt ggtcaggcca ttcacccttg ggctcgatgg ctgctcactg tcagagagca
     2641 caatcacatc aggcgaggga ggtctcttct cagacttcat gtcactgtgt gaagtgcgca
     2701 tgtccacagg cccgtctcct aacagcccct cgccagatcc tgtgcccatg gcctctgttg
     2761 tcctcagcaa cccctgggct gagcctgcac tcggctctgg catcaccctg ctgtctccgt
     2821 ccactgtcag ctctgatgac cctttctcca ttttcatttt cttgttctcc acatcatcct
     2881 ccgttaggtc aggttcaagg gtgcgcttct gactgcgggt tcggcatgct tcttcactgg
     2941 tgccaacttt tttgtacaaa cttgtcttgt catcgtcatc cttgtaatcg atgtcatgat
     3001 catatgtatc accgtcatgg tctttgtagt ccatggtgga agcttaagtt taaacgctag
     3061 aggatccagc ctccgcggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag
     3121 cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct tatatagggg
     3181 gtaccttctc tatcactgat agggagctcg agacgtgtcg acttctctat cactgatagg
     3241 gagacgttct agacgtacct tctctatcac tgatagggag ctcgagacgt gtcgacttct
     3301 ctatcactga tagggagacg tgcggccagc ttggatccct cgagacgcgt atatctggcc
     3361 cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg
     3421 gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac
     3481 acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     3541 gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc acttttcggg
     3601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     5641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     5701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag
     5761 aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5821 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5881 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
     5941 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     6001 aggcggcggt ggaatcgaaa tctcgtagta cgtgctattc ctttgccctc ggacgagtgc
     6061 tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg
     6121 cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt
     6181 cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga
     6241 gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg
     6301 gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga
     6361 agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga
     6421 ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga
     6481 ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga
     6541 cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa
     6601 tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tc
//
