LOCUS       sequence-311887-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Splicing reporter backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  AUTHORS   Adamson SI, Zhan L, Graveley BR
  TITLE     Vex-seq: high-throughput identification of the impact of genetic 
            variation on pre-mRNA splicing efficiency.
  JOURNAL   Genome Biol. 2018 Jun 1;19(1):71. doi: 10.1186/s13059-018-1437-x.
  PUBMED    29859120
REFERENCE   2  (bases 1 to 6955)
  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"; journalName: "Genome 
            Biol."; date: "2018-06-1"; volume: "19(1):71. doi"; pages: " 
            10.1186/s13059-018-1437-x"
COMMENT     Sequence Label: modified pcAT7-Glo1
FEATURES             Location/Qualifiers
     source          1..6955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        71..348
                     /label=chicken beta-actin promoter
     primer_bind     1289..1311
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1338..1357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1338..1356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2974..2991)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2980..3204
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3250..3678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3337..3356)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3547..3568
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3687..3707)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3692..4022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3873..4008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3935..3954
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4070..4117
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4136..4510
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4640..4761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4677..4696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4731..4750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4810..4826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4810..4826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4823..4845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4834..4850
                     /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)."
     promoter        complement(join(4858..4864,4865..4882,4883..4888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4903..4924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5041..5058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5212..5800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5292..5311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5971..6762,6763..6831))
                     /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     6594..6613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6832..6936)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatccc agtgtggtgg tacgtaggaa ttaattcgcc
       61 agcacagtgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      121 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      181 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      241 cggaaaggtg cggcggcagc caatcaaagc ggcgcgctcc gaaagtttcc ttttatggcg
      301 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      361 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      421 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      481 tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag ccttgagggg
      541 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      601 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
      661 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
      721 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
      781 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
      841 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
      901 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
      961 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     1021 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     1081 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     1141 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1201 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     1261 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1321 agcgtttaaa cttaagctaa tacgactcac tatagggagc ttggtaccgc aacctcaaac
     1381 agacaccatg gtgcacctga ctcctgagga gaagtctgcc gttactgccc tgtggggcaa
     1441 ggtgaacgtg gatgaagttg gtggtgaggc cctgggcagg ttggtatcaa ggttacaaga
     1501 caggtttaag gagaccaata gaaactgggc atatggagac agagaagact cttgggtttc
     1561 tgatagggcc cactgactct ctctgcctct gcaggccctg ggcaggttgg tatcaaggtt
     1621 acaagacagg tttaaggaga ccaatagatc tgggcatgtg gagacagaga agactcttgg
     1681 gtttctgata ggcactgact ctctctgcct attggtctat tttcccaccc ttaggctgct
     1741 ggtggtctac ccttggaccc agaggttctt tgagtccttt ggggatctgt ccactcctga
     1801 tgctgttatg ggcaacccta aggtgaaggc tcatggcaag aaagtgctcg gtgcctttag
     1861 tgatggcctg gctcacctgg acaacctcaa gggcaccttt gccacactga gtgagctgca
     1921 ctgtgacaag ctgcacgtgg atcctgagaa cttcagggtg agtctatggg acccttgatg
     1981 ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag
     2041 ggtacagttt agaatgggaa acagacgaat gattgcatca gtgtggaagt ctcaggatcg
     2101 ttttagtttc ttttatttgc tgttcataac aattgttttc ttttgtttaa ttcttgcttt
     2161 cttttttttt cttctccgca atttttacta ttatacttaa tgccttaaca ttgtgtataa
     2221 caaaaggaaa tatctctgag atacattaag taacttaaaa aaaaacttta cacagtctgc
     2281 ctagtacatt actatttgga atatatgtgt gcttatttgc atattcataa tctccctact
     2341 ttattttctt ttatttttaa ttgatacata atcattatac atatttatgg gttaaagtgt
     2401 aatgttttaa tatcgataca catattgacc aaatcagggt aattttgcat ttgtaatttt
     2461 aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct
     2521 aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa
     2581 agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt
     2641 ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca
     2701 gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct
     2761 aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg
     2821 tgctggtctg tgtgctggcc catcactttg gcaaagaatt caccccacca gtgcaggctg
     2881 cctatcagaa agtggtggct ggtgtggcta atgccctggc ccacaagtat cactaagctc
     2941 gctctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3001 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3061 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3121 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3181 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3241 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3301 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3361 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3421 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3481 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3541 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3601 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     3661 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3721 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3841 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3901 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3961 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4021 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     4081 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     4141 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     4201 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     4261 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     4321 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     4381 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     4441 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     4501 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4561 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4621 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4741 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4801 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4861 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4921 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4981 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5401 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5641 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtg
//
