LOCUS       sequence-421030-        5502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Chtop+exon(138nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5502)
  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 5502)
  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 Chtop+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(993..1040)
                     /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     1172..1191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1324..1548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1537..1554
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    1596..1620
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2371..2395)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2396..2419)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2571..2595)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2617..2635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2654..2672
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2691..2709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2728..2746
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2945..2964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3039..3143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3144..3212,3213..4004)
                     /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(3362..3381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4175..4763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4664..4683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4917..4934
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5051..5072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5087..5092,5093..5110,5111..5117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5125..5141
                     /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     5130..5152
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5149..5165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5149..5165
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5214..5335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5225..5244)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5279..5298
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5465..5502,1..985))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg
       61 cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct
      121 gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac
      181 caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct
      241 cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg
      301 cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc
      361 ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt
      421 cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga
      481 cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga
      541 aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg
      601 tctggctaag atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag
      661 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
      721 aataggtcag gctctcgctg aactccccaa tgtcaagcac ttccggaatc gggagcgcgg
      781 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
      841 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
      901 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
      961 acgtcgcggt gagttcaggc tttttggcca aggaagttcc tatactttct agagaatagg
     1021 aacttcggaa taggaacttc taggtacgtg aaccatcacc ctaatcaagt tttttggggt
     1081 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1141 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1201 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1261 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag
     1321 aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
     1381 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     1441 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     1501 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
     1561 agcgggttta aacgggccct ctagactcga gtcagaccac tttgtacaag aaagttgggt
     1621 atcaatcatt ggtttcagga tctgtctgtg ccatgtaggc atccaattca gcatccaggt
     1681 gtcctttagt tttcgacatg tatgcatcca attggttgtc cagctgctcc ttggtcaata
     1741 cagggcgagt gagggcacct ctccctcggc cacgacctcg gcctctgcct ccaaagcccc
     1801 ctcttccccg acctatcata ccccgacctc taccaccgat tccgccacga cccatagctc
     1861 cacgccctag gccccctctc ccaggacctc cacgacctcg aacaccacct cttcttaagc
     1921 ccattcgggg agctacggct cgtccacctc ggagcaggtt ttgacctcgg agcgacatcc
     1981 cacccctaag cagggttctg gtagcacgtc ccccacgtag tcctcctcgg ggcaagcctc
     2041 tctggattat gggtaggcct cttcctccaa ttgctcccct ggccagggca cctatgggtc
     2101 ggcctaaccg tgcctggata ttactcttac ccaggcgctg ctttaagctc ttctgcttaa
     2161 gttttaatgc tgcctggaca gagggtctat tctccatctg ctgggccagt cttctgtttc
     2221 tggcactggc tagctgctgc tgctgctgca tcgaagcccg aatattcact ggcatcggct
     2281 gtttgttctt cagcatatta gtaaagcgct catttagaga catcttggtg gtgcttttta
     2341 gcacaacttt cggcgctgac tgtgcagcgg tgccaacttt tttgtacaaa cttgtcttgt
     2401 catcgtcatc cttgtaatcg atgtcatgat catatgtatc accgtcatgg tctttgtagt
     2461 ccatggtgga agcttaagtt taaacgctag aggatccagc ctccgcggag gctggatcgg
     2521 tcccggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg
     2581 ttcactaaac gagctctgct tatatagggg gtaccttctc tatcactgat agggagctcg
     2641 agacgtgtcg acttctctat cactgatagg gagacgttct agacgtacct tctctatcac
     2701 tgatagggag ctcgagacgt gtcgacttct ctatcactga tagggagacg tgcggccagc
     2761 ttggatccct cgagacgcgt atatctggcc cgtacatcgc gaagcagcgc aaaacgccta
     2821 accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt
     2881 ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac
     2941 tatgcggcat cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg
     3001 atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3061 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4081 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4141 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4201 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4261 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4321 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4381 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4501 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4561 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4621 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4681 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4741 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4801 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4861 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4921 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     4981 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5041 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5101 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5161 atgaccatga ttacgccaag ctctagctag aggtcgacgg tatacagaca tgataagata
     5221 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     5281 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt
     5341 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     5401 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtagta
     5461 cgtgctattc ctttgccctc ggacgagtgc tggggcgtcg gt
//
