LOCUS       sequence-321881-        6954 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 6954)
  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 6954)
  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..6954
                     /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..4021
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3872..4007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3934..3953
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4069..4116
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4135..4509
                     /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    4639..4760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4676..4695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4730..4749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4809..4825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4809..4825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4822..4844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4833..4849
                     /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(4857..4863,4864..4881,4882..4887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4902..4923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5040..5057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5211..5799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5291..5310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5970..6761,6762..6830))
                     /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     6593..6612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6831..6935)
                     /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 aggggggggg ggggggggcg gggggggggg cggggcgagg
      241 cggagaggtg cggcggcagc caatcagagc 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 gcttgtttct 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 tcgggggctc 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 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3661 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3721 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3781 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3841 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3901 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3961 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4021 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     4081 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     4141 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     4201 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     4261 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     4321 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     4381 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     4441 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     4501 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4561 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4621 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4681 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4741 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4801 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4861 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4921 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4981 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5041 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5101 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5401 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5641 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5881 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5941 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6001 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6061 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6121 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6181 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6241 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6301 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6361 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6421 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6481 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6541 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6601 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6661 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6721 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6781 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6841 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6901 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtg
//
