LOCUS       sequence-150808-        5724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minigene reporter for identification and analysis of cis elements 
            and trans factors affecting pre-mRNA splicing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5724)
  AUTHORS   Singh G, Cooper TA
  TITLE     Minigene reporter for identification and analysis of cis elements 
            and trans factors affecting pre-mRNA splicing.
  JOURNAL   Biotechniques. 2006 Aug;41(2):177-81.
  PUBMED    16925019
REFERENCE   2  (bases 1 to 5724)
  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: 
            "Biotechniques."; date: "2006-08"; volume: "41(2)"; pages: "177-81"
COMMENT     Sequence Label: RHCglo
FEATURES             Location/Qualifiers
     source          1..5724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..70)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        347..608
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2881..2900)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2882..2900)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2907..2924)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2907..2923)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2916..2938)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3065..3520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3152..3171)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3362..3383
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3547..3651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3652..3720,3721..4512)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3870..3889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4683..5271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5425..5442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(5595..5600,5601..5618,5619..5625)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5633..5649
                     /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     5638..5660
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5657..5673
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5657..5673
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5694..5712
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtacatatc gaattcgagc tcgcccgggg atcatatggt gcactctcag tacaatctgc
       61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      241 atctgagggg actagggtgt gtttaggcga aaagcggggc ttcggttgta cgcggttagg
      301 agtcccctca ggatatagta gtttcgcttt tgcataggga gggggaaatg tagtcttatg
      361 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
      421 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
      481 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattccgc attgcagaga
      541 tattgtattt aagtgcctag ctcgatacaa taaacgccat ttgaccattc accacattgg
      601 tgtgcacctc caagctccgg actcgggtga gcccttatgc atgtctctta ctgaccagct
      661 tgctcttacc ggggttgcag ggaggcagat ttgctcacct gaatgttggg gaggtaaatg
      721 cttggagaga tgggggtgct gataaatcca tctctgtcct atgccggtca gagggtctgt
      781 gcatcgggag ctctccctct tcctttccca gctgccttct ttgcctttgg tttcatcctc
      841 acttggtttt atgctctgcc ttcgtctccc taactcctcc ctagtaggaa aaagcatcct
      901 ccgctgtggc accttgcaat tctccccatc tgcacccctc cctctgcctg ctgcgtgcag
      961 ccccttcctc cccaggttgc ccctcagcac ctgagtttcc gtaactcaag gactggagcc
     1021 cctgttactg accatacccc gctatggggc tgatctggca ttgctccccc acgccttttg
     1081 cgctttcagt gagggtgcac ggatatggga acgccagcaa aaatagctcc tgctcctcat
     1141 ccacctgtgg tggcttttgc ctcgcgtcgc cccaacctcc ctgagtcacg gggcctggct
     1201 gcgtctgagg agacagctgc agctccttgt gcagctcccc agccattttt agaagcactt
     1261 tccccacccc cctccccccc cctcttccca gcaatgtgtt gtgcccgcac attttccagg
     1321 ataaggtttc ctcggggagc ttggccctgc agcctttacc tcctcctcag gcaacgaccc
     1381 cgtcgatcct cactttgtag ttcctctgta ggcaaagcag ttgtggagtg gtgggaggaa
     1441 agaatgtccc atgacttggc tttcctgatg ccttctggag aagaaaggaa gggcgagcaa
     1501 aatccctttc gcgaagggcc tttgctggca gttggatgct ttcccccttt cctgcttccc
     1561 tagccggcca cgagatggtg ctgggaagcg aaagggtcga caccaataga aactgggcat
     1621 gtggagacag agaagactct tgggtttctg ataggcactg actctctctg cctattggtc
     1681 tattttccca cccttaggat ccatctacca tcgattggca tggctcgagg ttggtatcaa
     1741 ggttacaaga caggtttaag gagaccaata gaaactgggc atgtggagac agagaagact
     1801 cttgggtcta gactagtcga gctgcccgaa caacttcagg gccgcatctc atttgccagc
     1861 aggggggatg caaaaaatcc cacctatggt gctgatgttc aatgaatgca ttgggtgccc
     1921 actagcccag tgacccaggg aaggctagga aagcactccg agacaaagta gcacttttca
     1981 gaaggagaat aagccctggg cttggagtcc ttgaggaatc tgagcaagat tcctctggtt
     2041 aatggtgcac aatatctcta agctagaagc tgaagctggt ttggggacca aatgtgtgac
     2101 aagctgtggg attgtgtggg gaagaagaaa agatgaagga atgcatcgta acttaccttg
     2161 ttctcatttt tccctcctac agaaaaagag gagggcagcc accgcccggc ggcagcacct
     2221 gaagagtgct atgctccagc aatcttgtag gttccaggct gctgaggacc tgcaccagcc
     2281 atgcaacttt ctattttgta acaatttctg gttactgttg ctgcaaagct ccatgtgaca
     2341 cagtgtatgt aaagtgtaca taaattaatt tattttacct cgttttgttt gtttttaaaa
     2401 ccaatgccct gtggaaggaa acataaaact tcaagaagca ttaaatcatc agtcattctg
     2461 tcacacccct aatgcagttg tttctgtcat catttccctg ggctcttcca tctctcgctg
     2521 acctgggact gggtgctggg gctgggagca ggggttgggg ctctccaggg agagatggca
     2581 tggggagagt gatgggatac tgctgggggg gggggactca ccctgctgtg ggctgcagga
     2641 agcccattgg tgcagagagc agcctgggat gcccatgaca cgggcaccca ctgcaccgtg
     2701 tttctcccat gcccagtagg gaaagggtta cgagcgccgt tcattctcag cttgtgaagg
     2761 attttgttgg gctcagcctg ccagagcagt agccaggcat gcctgtgcag ctccgagctg
     2821 tgatggacag aggcaaggct gcagctagag cggccgccac cgcggtggag ctccaattcg
     2881 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2941 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccccttcgc cagctggcgt
     3001 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgtagcct gaatggcgaa
     3061 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3121 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3241 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3301 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3361 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3421 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3481 tttaacgcga attttaacaa aatattaacg tttacaattt caggtggcac ttttcgggga
     3541 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3601 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3661 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3721 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3781 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3841 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3901 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3961 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4021 gccataagca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4081 aaggagctaa ccgctttttt tcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4141 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4201 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4261 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4321 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4381 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggc
     4441 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4501 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4801 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5041 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     5101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5161 gagcttccag gggggaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5221 cttgagcgtc gatttttgtg atgctcgtca ggggggccga gcctatggaa aaacgccagc
     5281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5341 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5401 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5461 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5521 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta cctcactcat
     5581 taggcacccc aggctttaca ctttatgctt ccggctccta tgttgtgtgg aattgtgagc
     5641 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cggaattaac
     5701 cctcactaaa gggaacaaaa gctg
//
