LOCUS       sequence-240737-        6920 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 6920)
  AUTHORS   Kishore S, Khanna A, Stamm S
  TITLE     Rapid generation of splicing reporters with pSpliceExpress.
  JOURNAL   Gene. 2008 Dec 31;427(1-2):104-10. Epub 2008 Oct 1.
  PUBMED    18930792
REFERENCE   2  (bases 1 to 6920)
  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: "Gene."; 
            date: "2008-12-31"; volume: "427(1-2)"; pages: "104-10. Epub 2008 
            Oct 1"
COMMENT     Sequence Label: pSpliceExpress
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..112)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     161..183
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     175..192
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     176..192
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    209..440
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(836..1141)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(938..957)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1461..2144)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     2059..2078
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2145..2247)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2392..2623)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2641..2660)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2642..2660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2665..2681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2665..2681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3302..3320)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3388..3492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3493..3561,3562..4353)
                     /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(3711..3730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     4889..4908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(5463..5659)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5477..5612
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5767..5786
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      complement(6827..6854)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
       61 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
      121 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
      181 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
      241 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      301 gctgaatgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
      361 aaacagacta catcatactg taaaacacaa catatccagt cactatgaat caactactta
      421 gatggtatta gtgacctgta gtcgaccgac agccttccaa atgttcttcg ggtgatgctg
      481 ccaacttagt cgaccgacag ccttccaaat gttcttctca aacggaatcg tcgtatccag
      541 cctactcgct attgtcctca atgccgtatt aaatcataaa aagaaataag aaaaagaggt
      601 gcgagcctct tttttgtgtg acaaaataaa aacatctacc tattcatata cgctagtgtc
      661 atagtcctga aaatcatctg catcaagaac aatttcacaa ctcttatact tttctcttac
      721 aagtcgttcg gcttcatctg gattttcagc ctctatactt actaaacgtg ataaagtttc
      781 tgtaatttct actgtatcga cctgcagact ggctgtgtat aagggagcct gacatttata
      841 ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag
      901 ccacttcttc cccgataacg gagaccggca cactggccat atcggtggtc atcatgcgcc
      961 agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta tctgacagca
     1021 gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata atatcactct
     1081 gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc ttaaactgca
     1141 tttcaccagc ccctgttctc gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc
     1201 tcagccatcc cttcctgatt ttccgctttc cagcgttcgg cacgcagacg acgggcttca
     1261 ttctgcatgg ttgtgcttac cagaccggag atattgacat catatatgcc ttgagcaact
     1321 gatagctgtc gctgtcaact gtcactgtaa tacgctgctt catagcatac ctctttttga
     1381 catacttcgg gtatacatat cagtatatat tcttataccg caaaaatcag cgcgcaaata
     1441 cgcatactgt tatctggctt ttagtaagcc ggatccacgc ggcgtttacg ccccccctgc
     1501 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     1561 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     1621 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     1681 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     1741 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     1801 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     1861 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     1921 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     1981 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     2041 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     2101 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     2161 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     2221 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     2281 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     2341 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttagtcga ctacaggtca
     2401 ctaataccat ctaagtagtt gattcatagt gactggatat gttgtgtttt acagtattat
     2461 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     2521 tctcgttcag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2581 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2641 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctc tagagcggcc
     2701 gccaccgcgg tggagctcgg tacctatttg gggaccccat agagcactgc actgactgag
     2761 ggatggtaac aggatgtgta ggttttggag gcccatatgt ccattcatga ccagtgactt
     2821 gtctcacagc catgcaaccc ttgcctcctg tgctgactta gcaggggata aagtgagaga
     2881 aagcctgggc taatcagggg gtcgctcagc tcctcctaac tggattgtcc tatgtgtctt
     2941 tgcttctgtg ctgctgatgc tctgccctgt gctgacatga cctccctggc agtggcacaa
     3001 ctggagctgg gtggaggccc gtgaccttca gaccttggca ctggaggtgg cccggcagaa
     3061 gcgcggcatc gtggatcagt gctgcaccag catctgctct ctctaccaac tggagaacta
     3121 ctgcaactag gcccaccact accctgtcca cccctctgca atgaataaaa cctttgaaag
     3181 agcactacaa gttgtgtgta catgcgtgca tgtgcatatg tggtgcgggg ggaacatgag
     3241 tggggctggc tggagtggcg atgataagct gtcaaacatg agaattcttg aagacgaaag
     3301 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3361 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3481 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3541 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3601 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3661 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3721 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3781 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3841 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3901 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4021 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4081 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4141 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4321 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggctgc tgcttgcaaa
     4441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4981 tatggaaaaa cgccagcaac ggagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     5041 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     5101 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg
     5161 aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg
     5221 cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga
     5281 cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct
     5341 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     5401 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcggggagc
     5461 tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag
     5521 gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
     5581 tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt
     5641 gctgactaat tgagatgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     5701 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     5761 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     5821 ggccacgatg cgtccggcgt agaggatctc aggatatagt agtttcgctt ttgcataggg
     5881 agggggaaat gtagtcttat gcaatactct tgtagtcttg caacatgctt atgtaacgat
     5941 gagttagcaa catgccttat aaggagagaa aaagcaccgt gcatgccgat tggtgggagt
     6001 aaggtggtat gatcgtggta tgatcgtgcc ttgttaggaa ggcaacagac gggtctaaca
     6061 cggattggac gaaccactga attccgcatt gcagagatat tgtatttaag tgcctagctc
     6121 gatacaataa acgccatttg accattcacc acattggtgt gcacctcaag cttcctgcat
     6181 gctgctgctg ctgctgctgc tgggcctgag gctacagctc tccctgggca tcatcccagt
     6241 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     6301 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     6361 gggggtgtct acggtgacag ctgccaggat cgatccgctt cctgcccctg ctggccctgc
     6421 tcatcctctg ggagccccgc cctgcccagg cttttgtcaa acagcacctt tgtggttctc
     6481 acttggtgga agctctctac ctggtgtgtg gggagcgtgg attcttctac acacccatgt
     6541 cccgccgcga agtggaggac ccacaaggta agctctgctc ctgaattaat tctatcccaa
     6601 gtgctaacta ccctgtttgt ctttcaccct tgagaccttg taaattgtgc cctaggtgtg
     6661 gagggtctca ggctaaccag tggggggcac atttctgtgg gcagctagac atatgtaaac
     6721 atggtagctg ccaggaagga gtgagaatcc ttccttaagt ctcctaggtg gtgacgggtg
     6781 gctaggcccc aggataggta ccgggccccc cctcgagaga aacgcaaaaa ggccatccgt
     6841 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     6901 ccctccgggc cgttgcttca
//
