LOCUS       sequence-420999-        6912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LR vector for integration of Mta1-exon into N2a FRT rtTA3 expression
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6912)
  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 6912)
  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..6912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(288..335)
                     /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     467..486
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    619..843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     832..849
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     protein_bind    891..915
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3076..3100)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3101..3124)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3276..3300)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3322..3340
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3359..3377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3396..3414
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3433..3451
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3650..3669
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3744..3848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3849..3917,3918..4709)
                     /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(4067..4086)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4880..5468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5369..5388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5622..5639
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5756..5777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5792..5797,5798..5815,5816..5822)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5830..5846
                     /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     5835..5857
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5854..5870
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5854..5870
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5919..6040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5930..5949)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5984..6003
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6170..6912,1..280))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 cacggcggga gatgcaatag gtcaggctct cgctgaactc cccaatgtca agcacttccg
       61 gaatcgggag cgcggccgat gcaaagtgcc gataaacata acgatctttg tagaaaccat
      121 cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac
      181 gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac ttttcgatca
      241 gaaacttctc gacagacgtc gcggtgagtt caggcttttt ggccaaggaa gttcctatac
      301 tttctagaga ataggaactt cggaatagga acttctaggt acgtgaacca tcaccctaat
      361 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
      421 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
      481 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
      541 ccgccgcgct taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg
      601 ttctttccgc ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt
      661 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
      721 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
      781 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
      841 cagtcgaggc tgatcagcgg gtttaaacgg gccctctaga ctcgagtcag accactttgt
      901 acaagaaagt tgggtactag tcctcaataa caatgggctc atcattgact ggtgcaggtg
      961 ggggtggccg cagcggcagg gcctggctct ggcgcagggc aatgggtttg tagggccggc
     1021 gggcagcacg cttggtttct gaggatgaga gcagctttcg gattttcctg gtctcttcgg
     1081 tggccaagta aaatacatca tctggggcgt cgatccagtt cattcgccgc cgcttgactg
     1141 gaggcaggga gccaccacgg attaggcgca ctttggtggg gtaggacttc ccattgagca
     1201 ggagattccg acttggtccc atgtgcctgg tctgtccatg gtttgccaga cccaggtaag
     1261 tgcccctact gggcatcaag aggcgcttct tcatgttgcc atctacccca ttgtgctgct
     1321 cgtagctcct cttgcccagg atggtagggg agccgttgtt gatgacaggg gctgacttgg
     1381 caggggtcag actgctgagc acactggatg aactcttcac ggggtcgggc ttaggcgggc
     1441 ggggatgggt ctcaagatac cggagcacgg cctccagggg cttccgtact acctgcttca
     1501 gcaccagtgg gctctgggag gcttcgggca gccgtgctgt gcattcagcc ttgattgctg
     1561 cgctgttgat gggcatgtag gggtggcggg cagcatgcca tgggcgtagg atctcacggc
     1621 acaagcgccg ggcaatgcgt gtcaacttgg tagtatgcag gtagaaggcc tgccttgtct
     1681 tcatggcaaa cttggggctc ccactgctcc gagctgggat gccatgggga ctcatgttat
     1741 tgcggtttgg tcccggcctc tctccatcta accgggttgg cattttcaag ccaccatatt
     1801 tcttccaata tgtccaacaa gatgcgcaga ggcgacactg catgttaggg ggaccccaag
     1861 aataccactg gtaagactgt gtggtgtaac agctctcgca ggcccgaccg gccccggggc
     1921 tctggcctgg tgtccctgtg ccattcacta cagtggcctt gacactgctg gcgctgatct
     1981 ggtttgggtt gggcttgtta tagttgggaa tataaacctg ctttaacttg ctctcagctt
     2041 cagctgcttt caagcgtttc tgctggacgt atctgtcggt ggtcttccac atgtagtaat
     2101 actcaatgat gctggtgagc gacttccatg ggagaaaatc ttgttgaatg tcggtgaaat
     2161 ctttcccata tttttccaga gcttcctcaa aaaggttggc ttctgatgcg gaccactcct
     2221 ccatctcatc gcggcagagc actggcccac cctggggcac gagggccgag atggccttgg
     2281 agatgtcgta gatgttcttg tgcagcgtgt ccatagcatg gaacagggtg atgtctcggg
     2341 aggccgctgc ggcactcatg tgcaggctag gctgccgcac ggagctgctg caatccaggg
     2401 cccgtgcaaa ggtacccaca gagcgggcca ccaccagaaa ctggtcaatc tgcttatcca
     2461 caagtggatt gtgggcttcc cacaccttgg tctccagttt tgactgatcg cggccgtcct
     2521 cctcaccttc tttcagcaag tcagtgatgt cagcctggta ccggtttcct acacgaattt
     2581 cccctttatc agccaggagg gtcttctgct gtgggtcgta gactagagag tagaagaaga
     2641 aatcctcacg ctccaagtag gacttgagtg actccgtctc attgagcagg gtaacactgc
     2701 acttgcccct gatgtgggtg gcaggcagag actccaactg ccgagacagg aacagttctc
     2761 gatgccgcag ctgatgctta agtttctcag gcaggtccac catctctggg ttttctacct
     2821 cctcctccac ttccccttcc tcgccggtgt ccgcccccgg tccggctcta tagcagactg
     2881 acagggttgc atgcttgtcc gccagcgcga tgaggctgct ggaaatgtcc cgccttctgt
     2941 agaaacacac cactttggcc tccacattcc cgttggctgt cttgttaagc tcttcaatcc
     3001 tgcggatcag gtacgggttg ctggaggagt tctcaaagta gacatagtct ccgaccctgt
     3061 acatgttggc ggcggtgcca acttttttgt acaaacttgt cttgtcatcg tcatccttgt
     3121 aatcgatgtc atgatcatat gtatcaccgt catggtcttt gtagtccatg gtggaagctt
     3181 aagtttaaac gctagaggat ccagcctccg cggaggctgg atcggtcccg gtgtcttcta
     3241 tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct
     3301 ctgcttatat agggggtacc ttctctatca ctgataggga gctcgagacg tgtcgacttc
     3361 tctatcactg atagggagac gttctagacg taccttctct atcactgata gggagctcga
     3421 gacgtgtcga cttctctatc actgataggg agacgtgcgg ccagcttgga tccctcgaga
     3481 cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc
     3541 ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac
     3601 tcagcgacct ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag
     3661 cagattgtac tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag
     3721 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3781 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3841 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3901 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3961 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4021 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4081 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4141 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4201 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4261 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4321 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4381 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4441 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4501 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4561 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4621 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4681 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4741 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4801 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4861 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4921 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4981 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5041 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5101 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5161 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5221 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5281 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5341 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5401 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5461 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5521 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5581 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5641 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5701 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5761 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     5821 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     5881 ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg
     5941 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6001 tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca
     6061 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     6121 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tagtacgtgc tattcctttg
     6181 ccctcggacg agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat
     6241 cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg
     6301 atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa
     6361 ccaagctctg atagagttgg tcaagaccaa tgcggagcat atacgcccgg agccgcggcg
     6421 atcctgcaag ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca
     6481 accacggcct ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct
     6541 cgctccagtc aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga
     6601 aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat
     6661 cgagagcctg cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca
     6721 catggggatc agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt
     6781 gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca
     6841 tggcctccgc gaccggctgc agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg
     6901 tgacaccctg tg
//
