LOCUS       sequence-421016-        8424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Gse1+ex(108nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8424)
  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 8424)
  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 Gse1+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..8424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    46..64
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     263..282
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        357..461
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(462..530,531..1322)
                     /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(680..699)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1493..2081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1982..2001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2235..2252
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2369..2390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2405..2410,2411..2428,2429..2435)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2443..2459
                     /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     2448..2470
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2467..2483
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2467..2483
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2532..2653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2543..2562)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2597..2616
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2783..3805)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(3813..3860)
                     /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     3992..4011
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4144..4368
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4357..4374
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    4416..4440
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(6118..6138)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(8113..8137)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(8138..8161)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8313..8337)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    8359..8377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8396..8414
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 acgtaccttc tctatcactg atagggagct cgagacgtgt cgacttctct atcactgata
       61 gggagacgtg cggccagctt ggatccctcg agacgcgtat atctggcccg tacatcgcga
      121 agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc
      181 cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg
      241 agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg
      301 ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg
      361 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      421 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      481 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      541 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      601 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      661 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      721 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      781 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      841 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
      901 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
      961 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1021 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1081 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1141 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1201 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1261 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1321 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1381 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1441 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1501 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1561 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1621 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1681 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1741 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1801 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1861 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1921 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1981 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2041 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2101 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2161 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2221 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     2281 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2341 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     2401 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     2461 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta
     2521 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2581 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2641 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     2701 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     2761 aatcgaaatc tcgtagtacg tgctattcct ttgccctcgg acgagtgctg gggcgtcggt
     2821 ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg
     2881 cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct
     2941 gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac
     3001 caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct
     3061 cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg
     3121 cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc
     3181 ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt
     3241 cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga
     3301 cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga
     3361 aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg
     3421 tctggctaag atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag
     3481 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
     3541 aataggtcag gctctcgctg aactccccaa tgtcaagcac ttccggaatc gggagcgcgg
     3601 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
     3661 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
     3721 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
     3781 acgtcgcggt gagttcaggc tttttggcca aggaagttcc tatactttct agagaatagg
     3841 aacttcggaa taggaacttc taggtacgtg aaccatcacc ctaatcaagt tttttggggt
     3901 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3961 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4021 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4081 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag
     4141 aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
     4201 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     4261 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     4321 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
     4381 agcgggttta aacgggccct ctagactcga gtcagaccac tttgtacaag aaagttgggt
     4441 atcacctcgg atatccctta aagtagcccc taggccagtg catggtgggc aaggcaaggc
     4501 acttccgcaa gtggtccagc tccgcctgga gccgctcccg ctcagagacc atcttctgtt
     4561 tctgactccg aagctccgcc atgctgtgag acagctgctc cgcagtcagg ctgaggctgt
     4621 agttctgcgc ctccaaccgc cggcactgcg tctgcagcac ctgccgctcc aggttttgct
     4681 cttctatgtg ttcctggtaa gcttcaaaga ttgcctcaat cccttgccac tggcgcctcg
     4741 gggcttcctc agcttcctcc tcgctgtcct cctcagagtc ctcctcagag tcctggtccg
     4801 cctcctgcat aggagggccc ttcctcccca ttggaggctc ctgttgtcca ttgtgctggg
     4861 gcaaggggac cctgctggag gactgcagct cgggaatgtt gtagtggatt gccgtgtcga
     4921 ctttgtggct ctgctctgca gatagcaaag cagagttccc tttatgcttc tgcagagcct
     4981 tctgtgtgga ctgcagcacg gactcgtgga attgatgtgc aaactcttcg gccacgaaag
     5041 gctcccaggg cttgctcttc ccgttgatgc tgtgagacaa cggcacagga atgtccttgg
     5101 gcgcggcgcc ccggaggtaa tggagcatgc tcaggctctt cttgccacca ggggcctcgc
     5161 tcagcctggc cttctcacca ctgggaggag ccggctcctg gactctcaag agctcctggg
     5221 gccggagctg ttccagtctc ccggtctccg tggtctttgg gacatctgac aaggaggcag
     5281 ggaccccgac aggctgatcc gtctctagag gagctggctg tgtggctggt tcactcaggg
     5341 agacgggagg actgtccctc gaagagtttg tcactgagtc tgctgcggac agtgctctct
     5401 gcttcatggc tcggagcatt tcaacaagcc tctctttgtc tttcctcttc tccgcactga
     5461 tgtgcgtcaa gttgaagaag gtcaggaact tcctcttctc ctcaaagttg gggctgttgt
     5521 tcatctcgtc ggggctgtag cgggtggaca gagccagcct gggggaaggc gtctgtcgct
     5581 tgcactgaac ggcaggtggc gacgggcttc tctctctgag catccgcctc cgcttccggc
     5641 gcttctgcgc caccagttcc tccttctgct gttgggtggt caagccaaaa agttgcaaga
     5701 actctagctt ctccgaggat gtgtccagtt tgaggggcgg ctgctcagcc acacagcgga
     5761 ggtgtgccct gacctcctcc tcgtcactct cgtcgtacga gtcatcgagg tcgtagtagt
     5821 accctttctc ctgggcctcg cgccgcctgc gttcctccag gtccagtttg ctgaccagct
     5881 tccgtcgctg ctgaagaaac tcatcgtaga tgtaggctgg atcggggccc ctcgctgtgg
     5941 ggtggcagta cggagagcct ggtttcagag gcccactgag ctccccgaag gaggttgcct
     6001 gggaaagaga gggccgctgc tggcccaaga tggtctgtgt tgacttttca agttcagtaa
     6061 ggaaaggccc aggtccgtgg ggaaaggtct gaggctccag acttcctgcc tcccgcggtg
     6121 gtggtggtgg tggtggtggc tggtatttgt ccagtggtgt ggcctcacgc ttccggggct
     6181 cctctgcctt ctctgagcaa taaactcgct ccaccttcac cagtggcacg gcggcctggc
     6241 ggctgggctc aaaagcagtc gggtggctat gcagagagtg gctctcggcc cgccgcgtct
     6301 ccagagcatt gtccatcaga gacactgggt tccagagggc tgtgggtacg gtgtgctgct
     6361 ggggcttggg tgagatgagg ggtggagggc caccaaagtg ctgtgggggt tcacggctgc
     6421 cctgctcatg ccggtttgtt cctggcctgg tgctctcggg cctgtgctcc gcctctgtgg
     6481 gcaccagggg ccggcccagg tctaggtgct gctccaggac ctgctgccgg aactcggaca
     6541 cctgggactg gcggtcttcc ttctcctgcc gcaggcggcg ctgccgtgcc agccacttct
     6601 cctcctcgtt ggtgcgctgg atcagcaggg cggtggtggc gctgcttcca ggcagagaga
     6661 agatgccgtg gctggagatg aggctgggca ccgtgtggtg tggggcaggc actgggtgca
     6721 gagggtgctg cacgggcttg ggcgcctgca agcccacgtc cttcagcttc tctgctcggg
     6781 ttggggtcag ctgctccgag ggcttgggcc gctcctccgt gctgtgacct cggagtccgt
     6841 gcagctcggc cacaggcagg aaggtggtgg gctctaaggc cttggcagcc agcagctcct
     6901 tctccctcgc ccgctgctcc cgctgccgct ccagctcgcg ctcccgttcc ttctcccgct
     6961 cgcgctccag ctccttctcc cgctccttct cccgctgctc ccgctcccgc tccttctccc
     7021 gctcccggtc ggcctcccgt tcccgttccc gctctcgctc tctctcccgc cgcagttcct
     7081 cgtccatttg caacctctct gcactgaggc tggacatccg ctccgagtgt agcgccgcca
     7141 gggacgagtg ggagagctca ggaggatacc gtacgccgga gagatgcagg tgcatcgccg
     7201 acggatgcag tggaggcagg gagccagggg tggggatggg gtagaacgga gaccttaagg
     7261 ctgacaggca ataggagtca tccatcctga aggccgggtg ggggaatggg tgtggggcca
     7321 ggtagctggg gtgatagtag gctgcggcgg tggctgggtc caggccaagc ggaggcaagg
     7381 aggacatgcg taagtcttca gcagtgtggt agggccggaa gcttctcaag tagtcctcag
     7441 tcaccgtgct ggggggcacc acatggtgca caggccgctg gaggttgagt ggttggaagc
     7501 gggagtcctg tacaactgag ccgggggaaa ggccaaaggg gtacggggtg ctgagtaggt
     7561 gggaggggat ggcggggccc gcagcctttt cctgggcgag aggtggctcc accaataggc
     7621 gctcccgacc actgctgctg cctcgtgagc cagagtcctg gcggctctcg cttctccaga
     7681 ctccattgac ggttttagtg ggggcaatag tgaccactgg gggagtgctg gggacactgt
     7741 gacccccggg ggggacgatg atggggccca tgggcactcg tttgggcgtg ctagctggag
     7801 agctatggtt ggtggctgga gaggacactg gagatgactc gctgctcagt gaggaccctc
     7861 tgggctcctc tgcctgtttg gcgagcttgc gcagtgcggc agcaaagctg gaggatggag
     7921 cggcctgggc cgacagcgta ctgctggtgg ccgggctacc ggtgggcacc agggcgccat
     7981 tgagtggcga gggggtgagg gggttgacgg tggcggtggt cctggtcgcc gtggaaagca
     8041 tccctatgga aggggacttg ggctcatggc tcatgcctgg taagtaatag agaggcggct
     8101 tcagtccgaa ggtgccaact tttttgtaca aacttgtctt gtcatcgtca tccttgtaat
     8161 cgatgtcatg atcatatgta tcaccgtcat ggtctttgta gtccatggtg gaagcttaag
     8221 tttaaacgct agaggatcca gcctccgcgg aggctggatc ggtcccggtg tcttctatgg
     8281 aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg
     8341 cttatatagg gggtaccttc tctatcactg atagggagct cgagacgtgt cgacttctct
     8401 atcactgata gggagacgtt ctag
//
