LOCUS       sequence-37277-e        5730 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 5730)
  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 5730)
  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: pDESTsplice
FEATURES             Location/Qualifiers
     source          1..5730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1049..1065
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1050..1066
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1093..1217
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1242..1247,1248..1265,1266..1272)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1326..2006
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(1392..1411)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2326..2631
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2510..2529
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2672..2796)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2806..2822)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2806..2822)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3450..3468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3536..3640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3641..3709,3710..4501)
                     /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(3859..3878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5051..5070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaggatata gtagtttcgc ttttgcatag ggagggggaa atgtagtctt atgcaatact
       61 cttgtagtct tgcaacatgc ttatgtaacg atgagttagc aacatgcctt ataaggaaag
      121 aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt atgatcgtgg tatgatcgtg
      181 gtatgatcgt gccttattag gaaggcaaca gacgggtcta acacggattg gacgaaccac
      241 tgaattccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgccat
      301 ttgaccattc accacattgg tgtgcacctg ggttgatggc cggaccgttg attccctgac
      361 gactacgagc acatgcatga agcagaaggc ttcatttggt gacaagcttc tgcatgctgc
      421 tgctgctgct gctgctgggc ctgaggctac agctctccct gggcatcatc ccagttgagg
      481 aggagaaccc ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc
      541 tgcagcctgc acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg
      601 tgtctacggt gacagctgcc aggatcgatc cgcttcctgc ccctgctggc cctgctcatc
      661 ctctgggagc cccgccctgc ccaggctttt gtcaaacagc acctttgtgg ttctcacttg
      721 gtggaagctc tctacctggt gtgtggggag cgtggattct tctacacacc catgtcccgc
      781 cgcgaagtgg aggacccaca aggtaagctc tgctcctgaa ttaattccta tcccaagtgc
      841 taactaccct gtttgtcttt cacccttgag accttgtaaa ttgtgcccta ggtgtggagg
      901 gtctcaggct aaccagtggg gggcacattt ctgtgggcag ctagacatat gtaaacatgg
      961 tagctgccaa gaaggagtga gaatccttcc ttaagtctcc taggtggtga cgggtggcta
     1021 ggccccagga taggtaccgg gccccccctc gaggtcgacg gtatcgataa gctaattcct
     1081 gcagcccatc aaacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     1141 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     1201 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     1261 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     1321 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1381 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1441 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1501 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     1561 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1621 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1681 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1741 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1801 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1861 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     1921 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg caggcggggc
     1981 gtaatctaga ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga
     2041 tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat
     2101 gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca
     2161 tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc
     2221 tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta
     2281 ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt
     2341 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
     2401 gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
     2461 gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
     2521 accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac
     2581 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     2641 tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca
     2701 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
     2761 ttacgtttct cgttcagctt tcttgtacaa agtggttcga tgggggatcc actagttcta
     2821 gagcggccgc caccgcggtg gagctcggta cccatttggg gaccccatag agcaccgcac
     2881 cgaccgaggg atggtaacag gatgtgtagg ttttggaggc ccatatgtcc attcatgacc
     2941 agtgacttgt ctcacagcca tgcaaccctt gcctcctgtg ctgacttagc aggggataaa
     3001 gtgagagaaa gcctgggcta atcggggggt cgctcggctc ctcctaactg gattgtccta
     3061 tgtgtctttg cttctgtgct gctgatgctc tgccctgtgc tgacatgacc tccctggcag
     3121 tggcacaact ggagctgggt ggaggcccgt gaccttcaga ccttggcact ggaggtggcc
     3181 cggcagaagc gcggcatcgt ggatcagtgc tgcaccagca tctgctctct ctaccaactg
     3241 gagaactact gcaactaggc ccaccactac cctgtccacc cctctgcaat gaataaaacc
     3301 tttgaaagag cactacaagt tgtgtgtaca tgcgtgcgtg tgcatatgtg gtgcgggggg
     3361 aacatgagtg gggtcggctg gagtggtcgc ggcttaatct atctggcgat gataagctgt
     3421 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3481 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3541 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3601 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3661 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3721 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3781 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3841 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     3901 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3961 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4021 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4081 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4141 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     4201 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4261 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4321 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4381 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4441 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4501 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4561 taaaaggatc taggtgaaga tccgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4681 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4921 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5161 acgtcgggat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     5221 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     5281 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     5341 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     5401 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt
     5461 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     5521 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     5581 aagaatcata atggggaagg ccatccagcc tcgcgtcgag ctttttgcaa aagcctaggc
     5641 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     5701 ctgcataaat aaaaaaaatt agtcagccat
//
