LOCUS       sequence-420987-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mta1+exon(12nt) into N2a FRT rtTA3 
            expression cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  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 6900)
  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 Mta1+exon minimal CMV pCDNA5
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(294..341)
                     /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     473..492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    625..849
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     838..855
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    897..921
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3070..3094)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3095..3118)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3270..3294)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3316..3334
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3353..3371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3390..3408
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3427..3445
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3644..3663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3738..3842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3843..3911,3912..4703)
                     /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(4061..4080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4874..5462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5363..5382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5616..5633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5750..5771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5786..5791,5792..5809,5810..5816)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5824..5840
                     /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     5829..5851
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5848..5864
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5848..5864
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5913..6034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5924..5943)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5978..5997
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6164..6900,1..286))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 cctgtgcacg gcgggagatg caataggtca ggctctcgct gaactcccca atgtcaagca
       61 cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga tctttgtaga
      121 aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca tcgaagctga
      181 aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt
      241 cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttggcc aaggaagttc
      301 ctatactttc tagagaatag gaacttcgga ataggaactt ctaggtacgt gaaccatcac
      361 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
      421 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
      481 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
      541 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc
      601 agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca tgcctgctat
      661 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
      721 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
      781 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
      841 aaggcacagt cgaggctgat cagcgggttt aaacgggccc tctagactcg agtcagacca
      901 ctttgtacaa gaaagttggg tactagtcct caataacaat gggctcatca ttgactggtg
      961 caggtggggg tggccgcagc ggcagggcct ggctctggcg cagggcaatg ggtttgtagg
     1021 gccggcgggc agcacgcttg gtttctgagg atgagagcag ctttcggatt ttcctggtct
     1081 cttcggtggc caagtaaaat acatcatctg gggcgtcgat ccagttcatt cgccgccgct
     1141 tgactggagg cagggagcca ccacggatta ggcgcacttt ggtggggtag gacttcccat
     1201 tgagcaggag attccgactt ggtcccatgt gcctggtctg tccatggttt gccagacccc
     1261 tactgggcat caagaggcgc ttcttcatgt tgccatctac cccattgtgc tgctcgtagc
     1321 tcctcttgcc caggatggta ggggagccgt tgttgatgac aggggctgac ttggcagggg
     1381 tcagactgct gagcacactg gatgaactct tcacggggtc gggcttaggc gggcggggat
     1441 gggtctcaag ataccggagc acggcctcca ggggcttccg tactacctgc ttcagcacca
     1501 gtgggctctg ggaggcttcg ggcagccgtg ctgtgcattc agccttgatt gctgcgctgt
     1561 tgatgggcat gtaggggtgg cgggcagcat gccatgggcg taggatctca cggcacaagc
     1621 gccgggcaat gcgtgtcaac ttggtagtat gcaggtagaa ggcctgcctt gtcttcatgg
     1681 caaacttggg gctcccactg ctccgagctg ggatgccatg gggactcatg ttattgcggt
     1741 ttggtcccgg cctctctcca tctaaccggg ttggcatttt caagccacca tatttcttcc
     1801 aatatgtcca acaagatgcg cagaggcgac actgcatgtt agggggaccc caagaatacc
     1861 actggtaaga ctgtgtggtg taacagctct cgcaggcccg accggccccg gggctctggc
     1921 ctggtgtccc tgtgccattc actacagtgg ccttgacact gctggcgctg atctggtttg
     1981 ggttgggctt gttatagttg ggaatataaa cctgctttaa cttgctctca gcttcagctg
     2041 ctttcaagcg tttctgctgg acgtatctgt cggtggtctt ccacatgtag taatactcaa
     2101 tgatgctggt gagcgacttc catgggagaa aatcttgttg aatgtcggtg aaatctttcc
     2161 catatttttc cagagcttcc tcaaaaaggt tggcttctga tgcggaccac tcctccatct
     2221 catcgcggca gagcactggc ccaccctggg gcacgagggc cgagatggcc ttggagatgt
     2281 cgtagatgtt cttgtgcagc gtgtccatag catggaacag ggtgatgtct cgggaggccg
     2341 ctgcggcact catgtgcagg ctaggctgcc gcacggagct gctgcaatcc agggcccgtg
     2401 caaaggtacc cacagagcgg gccaccacca gaaactggtc aatctgctta tccacaagtg
     2461 gattgtgggc ttcccacacc ttggtctcca gttttgactg atcgcggccg tcctcctcac
     2521 cttctttcag caagtcagtg atgtcagcct ggtaccggtt tcctacacga atttcccctt
     2581 tatcagccag gagggtcttc tgctgtgggt cgtagactag agagtagaag aagaaatcct
     2641 cacgctccaa gtaggacttg agtgactccg tctcattgag cagggtaaca ctgcacttgc
     2701 ccctgatgtg ggtggcaggc agagactcca actgccgaga caggaacagt tctcgatgcc
     2761 gcagctgatg cttaagtttc tcaggcaggt ccaccatctc tgggttttct acctcctcct
     2821 ccacttcccc ttcctcgccg gtgtccgccc ccggtccggc tctatagcag actgacaggg
     2881 ttgcatgctt gtccgccagc gcgatgaggc tgctggaaat gtcccgcctt ctgtagaaac
     2941 acaccacttt ggcctccaca ttcccgttgg ctgtcttgtt aagctcttca atcctgcgga
     3001 tcaggtacgg gttgctggag gagttctcaa agtagacata gtctccgacc ctgtacatgt
     3061 tggcggcggt gccaactttt ttgtacaaac ttgtcttgtc atcgtcatcc ttgtaatcga
     3121 tgtcatgatc atatgtatca ccgtcatggt ctttgtagtc catggtggaa gcttaagttt
     3181 aaacgctaga ggatccagcc tccgcggagg ctggatcggt cccggtgtct tctatggagg
     3241 tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt
     3301 atataggggg taccttctct atcactgata gggagctcga gacgtgtcga cttctctatc
     3361 actgataggg agacgttcta gacgtacctt ctctatcact gatagggagc tcgagacgtg
     3421 tcgacttctc tatcactgat agggagacgt gcggccagct tggatccctc gagacgcgta
     3481 tatctggccc gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg
     3541 caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg
     3601 acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt
     3661 gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca
     3721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3901 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3961 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4021 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4081 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4141 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4201 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4261 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4321 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4381 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4441 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4501 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4561 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4621 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4681 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4741 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4801 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4861 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4921 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4981 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5041 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5101 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5161 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5221 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5281 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5341 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5401 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5461 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5521 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5581 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5641 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5701 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5761 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5821 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     5881 tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac
     5941 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     6001 atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga
     6061 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     6121 tttcatagaa ggcggcggtg gaatcgaaat ctcgtagtac gtgctattcc tttgccctcg
     6181 gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc
     6241 agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct ccggatcgga
     6301 cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc
     6361 tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg
     6421 caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg
     6481 gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc gcctcgctcc
     6541 agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag ccgaaatccg
     6601 cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc tcatcgagag
     6661 cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga tacacatggg
     6721 gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc
     6781 cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca tccatggcct
     6841 ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc aacgtgacac
//
