LOCUS       sequence-6777-ex        6238 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 6238)
  AUTHORS   Brun C, Dubey DD, Huberman JA
  TITLE     pDblet, a stable autonomously replicating shuttle vector for 
            Schizosaccharomyces pombe.
  JOURNAL   Gene. 1995 Oct 16. 164(1):173-7.
  PUBMED    7590311
REFERENCE   2  (bases 1 to 6238)
  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. 1995 
            Oct 16. 164(1):173-7."
COMMENT     Sequence Label: pDblet
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(625..1419)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     rep_origin      complement(2007..2462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2144..2165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2245..2823)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     2356..2375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2592..2614
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2606..2623
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2607..2623
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2633..2652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2633..2651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2660..2767
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2676..2692
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2677..2693
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2727..2743)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2727..2743)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2780..2800)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2780..2798)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2819..2835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2819..2835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2832..2854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2843..2859
                     /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(2867..2873,2874..2891,2892..2897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2912..2933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3050..3067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3221..3809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3301..3320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3980..4771,4772..4840))
                     /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     4603..4622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4841..4945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6211..6229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgga gcttgtgata ttgacgaaac tttttgacat ctaatttatt ctgttccaac
      241 accaatgttt ataaccaagt tttatcttgt ttgtctacat ggtattttac attcatctac
      301 atacatcttt cattggcttt gtacatagtt atcattacaa gtctaaaaaa attcactctt
      361 ttcttattca atgtcaatcc aagagaaaag attgtggtaa tgttgtagga gcatgtttaa
      421 taaattacta tagcaaatta ctttttattc ccaaggtgtt tatctataat agttaatatt
      481 ttagtcgcta cataaaattt taccaaagag tacttgtata ctaattctaa atgccttctg
      541 acataaaacg cctaggaaaa caaacgcaaa caaggcatcg actttttcaa taaccaacca
      601 aaaaaatttt acattagtct ttttttaatg ctgagaaagt ctttgctgat atgccttcca
      661 accagcttct ctatatctct tggcttcgac aacaggatta cgaccagctc catagactcc
      721 acgaccaaca atgatgatat cgctaccgca gtttacaatc acttcttcag gagtacgata
      781 ttgctgtccc agcccgtctc ctttaacatc caagccgata ccaggggaca tagttatgta
      841 gtcgctttga aggttaggaa atcgacgacc agctataaag ccaaagcaaa aatcggtatg
      901 cttctcaaac cattctaagg ttttctctgt gtaggaacca gtagccaaag agcctttgga
      961 agacatttca gccaaaagca agagaccacg tcccaaaggt aaaccaactt ctttgaggcc
     1021 ttgtataata ccctcgcctg gcactgtatg gcaatttgtg atatgagccc aagaagcaat
     1081 tttgtacaca ccagatgcat attgtagctt gacggtattt ccaatgtctg cgaatttgcg
     1141 atcctcaaag ataagaaaac gatgcttttt acctaaggcc accagttttt ctaccatatc
     1201 ctggtcgaaa tcctcgacaa cgtcaatatg tgtcttgata acacagacat agggtccaat
     1261 tttatctacc aattctaaga tttcggattt cttcgtcaaa tcgaccgcga ctgacaagtt
     1321 gctttgcttt tcttccatca aagccaacaa ttccttggca atgggatttt tcatcccctc
     1381 agctctagct gaatagcttt gaaatactct agcatccata actttgcttt taaaccttta
     1441 atttcgatcc aagcaaaaaa gaggttcttg gtaggacaat acggtaagaa aacacgacat
     1501 gtgcagagat gccgacgaag catagttaaa ctgggatggt aaaatcaatt aagaatttat
     1561 aaagacaaaa ttgtataagt ctctaaaaca tcttaattat acctcacaga actatctaaa
     1621 atatattcac aaagtgcaaa cattatcatg aaaaagaacc attttaattt aaagcaaggg
     1681 cattaaggct tatttacaga atttcttact tttgtaaaga ttataaggct gattatcttt
     1741 ttcaccatgc caaaaattac acaagataga atggatgttt gaaattaaac gtgagtatac
     1801 aaacaaatac actaggtaaa tcgaaacatt tttttctcca ttaagtaaca aattcctatt
     1861 tagagaaaga atgctgagta gattaaataa tctatacaaa cttttttaac acaaatgcat
     1921 acatatagcc agtgggattt gtagctaagc tccatatgcg gtgtgaaata ccgcacagat
     1981 gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt taaaattcgc
     2041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2101 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2221 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     2521 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     2581 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     2641 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct
     2701 tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt
     2761 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     2821 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2881 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2941 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3001 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3061 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3121 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3661 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3721 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3781 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3841 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3901 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3961 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4021 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4081 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4141 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4201 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4261 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4321 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4381 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4441 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4501 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4561 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4621 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4681 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4741 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4801 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4861 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4921 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcccta
     4981 gtggatcctt ctaggcgact aatacataaa ctcttttact tgataaattc aattttataa
     5041 ttgtgatgaa aagctaaagt tttactccaa attatataaa taaataattt ttgtaaattt
     5101 gatagatatt ttatgtatag gtatagatat atatatacta acaagggatt gtaataatat
     5161 tatataaaaa gtctattacc ttttgtatag cttacaatat tgtaacaaat taagaaataa
     5221 atcatataaa ataggaataa tttttagcga actctttgca ttagaacgaa tgaattttgt
     5281 atattgtata ttattcatga gacccatcta atactatttt ttatattaat taaaaaaaaa
     5341 aaaaaaaaaa acctgttgaa atatgtattt ggcgctaaac aatctctgaa tactaataaa
     5401 attttcgcta taaatattat cttgtcattg taatgtcatt tttttggctg tggaaatata
     5461 tcgtaaaagt cataacttat ccaaaagcta tcaaaaatgc aaaaaaatgc atcaaagttg
     5521 atacttagaa aacggcataa acatgccatc gtcttcattc ataaaattat ggatccttct
     5581 aggcgactaa tacataaact cttttacttg ataaattcaa ttttataatt gtgatgaaaa
     5641 gctaaagttt tactccaaat tatataaata aataattttt gtaaatttga tagatatttt
     5701 atgtataggt atagatatat atatactaac aagggattgt aataatatta tataaaaagt
     5761 ctattacctt ttgtatagct tacaatattg taacaaatta agaaataaat catataaaat
     5821 aggaataatt tttagcgaac tctttgcatt agaacgaatg aattttgtat attgtatatt
     5881 attcatgaga cccatctaat actatttttt atattaatta aaaaaaaaaa aaaaaaaaac
     5941 ctgttgaaat atgtatttgg cgctaaacaa tctctgaata ctaataaaat tttcgctata
     6001 aatattatct tgtcattgta atgtcatttt tttggctgtg gaaatatatc gtaaaagtca
     6061 taacttatcc aaaagctatc aaaaatgcaa aaaaatgcat caaagttgat acttagaaaa
     6121 cggcataaac atgccatcgt cttcattcat aaaattatgg atcccccgga cgtctaagaa
     6181 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
