LOCUS       sequence-345952-        8109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8109)
  AUTHORS   Min H, Turck CW, Nikolic JM, Black DL
  TITLE     A new regulatory protein, KSRP, mediates exon inclusion through an 
            intronic splicing enhancer.
  JOURNAL   Genes Dev. 1997 Apr 15. 11(8):1023-36.
  PUBMED    9136930
REFERENCE   2  (bases 1 to 8109)
  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: "Genes Dev. 
            1997 Apr 15. 11(8):1023-36."
COMMENT     Sequence Label: pET15b-KSRP
FEATURES             Location/Qualifiers
     source          1..8109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             97..114
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2611..2629)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2625..2672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2847..2852,2853..2869,2870..2875))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2902..2920)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2988..3092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3093..3161,3162..3953)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3311..3330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4124..4712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4613..4632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4866..4883
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4898..5037
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4963..4982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5123..5145
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5139..5330)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7685..7704
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7724..7801)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8007..8026
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagccatc atcaccatca tcacagcggc
      121 ggccacatgg tgtcggacta cagcacggga ggacccccgc ccgggccgcc gccgcccgcc
      181 ggcgggggcg ggggagccgg aggcgccggg ggaggccctc cgccgggccc gccaggcgcg
      241 ggggaccggg gcggcggcgg tcccggcggc ggcggcccgg gcggggggtc ggccgggggc
      301 ccctctcagc cacccggcgg aggcggcccg ggaatccgca aggacgcttt cgccgacgcc
      361 gtgcagcggg cccgccagat tgcagccaaa attggaggcg atgctgccac gacagtgaat
      421 aacagcactc ctgattttgg ttttgggggc caaaagagac agttggaaga tggagatcaa
      481 ccggagagca agaagctggc ttcccaggga gactcaatca gttctcaact tggacccatc
      541 catcctcccc caaggacttc aatgacagaa gagtacaggg tcccagacgg catggtgggc
      601 ctgatcattg gcagaggagg tgaacaaatt aacaaaatcc aacaggattc aggctgcaaa
      661 gtacagattt ctccagacag cggtggccta cccgagcgca gtgtgtcctt gacaggagcc
      721 ccagaatctg tccagaaagc caagatgatg ctggatgaca ttgtgtctcg gggtcgtggg
      781 ggccccccag gacagttcca cgacaacgcc aacgggggcc agaacggcac cgtgcaggag
      841 atcatgatcc ccgcgggcaa ggccggcctg gtcattggca agggcgggga gaccattaag
      901 cagctgcagg aacgcgctgg agtgaagatg atcttaattc aggacggatc tcagaatacg
      961 aatgtggaca aacctctccg catcattggg gatccttaca aagtgcagca agcctgtgag
     1021 atggtgatgg acatcctccg ggaacgtgac caaggcggct ttggggaccg gaatgagtac
     1081 ggatctcgga ttggcggagg catcgatgtg ccagtgccca ggcattctgt tggcgtggtc
     1141 attggccgga gtggagagat gatcaagaag atccagaatg atgctggcgt gcggatacag
     1201 ttcaagcaag atgacgggac agggcccgag aagattgctc atataatggg gcccccagac
     1261 aggtgcgagc acgcagcccg gatcatcaac gacctcctcc agagcctcag gagtggtccc
     1321 ccaggtcctc cagggggtcc aggcatgccc ccggggggcc gaggccgagg aagaggccaa
     1381 ggcaattggg gtccccctgg cggggagatg accttctcca tccccactca caagtgtggg
     1441 ctggtcatcg gccgaggtgg cgagaatgtg aaagccataa accagcagac gggagccttc
     1501 gtagagatct cccggcagct gccacccaac ggggacccca acttcaagtt gttcatcatc
     1561 cggggttcac cccagcagat tgaccacgcc aagcagctta tcgaggaaaa gatcgagggt
     1621 cctctctgcc cagttggacc aggcccaggt ggcccaggcc ctgctggccc aatggggccc
     1681 ttcaatcctg ggcccttcaa ccaggggcca cccggggctc ccccacatgc cggggggccc
     1741 cctcctcacc agtacccacc ccagggctgg ggcaatacct acccccagtg gcagccgcct
     1801 gctcctcatg acccaagcaa agcagctgca gcggccgcgg accccaacgc cgcgtgggcc
     1861 gcctactact cacactacta ccagcagccc ccgggccccg tccccggccc cgcaccggcc
     1921 cctgcggccc caccggctca gggtgagccc cctcagcccc cacccaccgg ccagtcggac
     1981 tacactaagg cctgggaaga gtattacaaa aagatcggcc agcagcccca gcagcccgga
     2041 gcgcccccac agcaggacta cacgaaggct tgggaggagt actacaagaa gcaagcgcaa
     2101 gtggccaccg gagggggtcc aggagctccc ccaggctccc agccagacta cagtgccgcc
     2161 tgggcggaat attacagaca gcaggccgct tactacggac agaccccagg tcctggcggc
     2221 ccccagccgc cgcccacgca gcagggacag cagcaggctc aatgaatcga atgaatgtga
     2281 acttcttcat ctgtgaaaaa tctttttttt ttccattttg ttctgtttgg gggcttctgt
     2341 tttgtttggc gagagagcga tggctgccgt ggggagtact ggggagccct cgcggcaagc
     2401 agggtggggg ggacttgggg gcatgccggg ccctcactct ctcgcctgtt ctgtgtctca
     2461 catgcttttt ctttcaaaat tgggatcctt ccatgttgag ccagccagag aagatagcga
     2521 gatctaaatc tctgccnaaa aaaaaaaaaa aaaaactcga ggatccggct gctaacaaag
     2581 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     2641 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     2701 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     2761 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     2821 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     2881 agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2941 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3001 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3061 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3121 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3181 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3241 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3301 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3361 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3421 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3481 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3541 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3601 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     3661 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3721 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3781 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3841 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3901 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3961 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4021 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4081 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4141 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4201 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4261 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4321 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4381 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4441 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4501 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4561 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4621 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4681 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4741 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4801 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4861 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4921 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     4981 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     5041 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5101 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5161 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     5221 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     5281 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     5341 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     5401 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     5461 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     5521 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     5581 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     5641 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     5701 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     5761 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     5821 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     5881 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     5941 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     6001 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     6061 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag
     6121 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     6181 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     6241 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     6301 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     6361 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     6421 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     6481 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     6541 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     6601 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     6661 cgggaaactg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6721 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     6781 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     6841 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     6901 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     6961 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     7021 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     7081 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     7141 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     7201 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     7261 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     7321 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     7381 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     7441 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     7501 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     7561 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     7621 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     7681 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     7741 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     7801 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     7861 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     7921 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     7981 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     8041 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     8101 ccgcgaaat
//
