LOCUS       sequence-43387-e        4328 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 4328)
  AUTHORS   Chee MK, Haase SB
  TITLE     New and Redesigned pRS Plasmid Shuttle Vectors for Genetic 
            Manipulation of Saccharomycescerevisiae.
  JOURNAL   G3 (Bethesda). 2012 May;2(5):515-26. Epub 2012 May 1.
  PUBMED    22670222
REFERENCE   2  (bases 1 to 4328)
  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: "G3 
            (Bethesda)."; date: "2012-05"; volume: "2(5)"; pages: "515-26. Epub 
            2012 May 1"
COMMENT     Sequence Label: pRS40B
FEATURES             Location/Qualifiers
     source          1..4328
                     /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"
     terminator      complement(249..446)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(839..1182)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     rep_origin      complement(1299..1754)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1436..1457)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1648..1667
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1731..2111)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1880..1902
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1894..1911
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1895..1911
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1921..1940
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1921..1939
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1948..2055
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1964..1980
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1965..1981
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2015..2031)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2015..2031)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2068..2088)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2068..2086)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2107..2123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2107..2123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2120..2142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2131..2147
                     /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(2155..2161,2162..2179,2180..2185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2200..2221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2338..2355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2509..3097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2589..2608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3268..4059,4060..4128))
                     /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     3891..3910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4129..4233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4301..4319
                     /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 accataatca tcgatgaatt cgagctcgtt ttcgacactg gatggcggcg ttagtatcga
      241 atcgacagca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc
      301 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
      361 acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
      421 aaaacaagaa tctttttatt gtcagtactg atcatgagat gcctgcaagc aattcgttct
      481 gtatcaggcg caggagcgtc ccgtccgggt cgaccaaagc ggccatcgtg cctccccact
      541 cctgcagttc gggggcatgg atgcgcggat agccgctgct ggtttcctgg atgccgacgg
      601 atttgcactg ccggtagaac tccgcgaggt cgtccagcct caggcagcag ctgaaccaac
      661 tcgcgagggg atcgagcccg gggtgggcga agaactccag catgagatcc ccgcgctgga
      721 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
      781 cggtgggatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atacccatgg
      841 ttgtttatgt tcggatgtga tgtgagaact gtatcctagc aagattttaa aaggaagtat
      901 atgaaagaag aacctcagtg gcaaatccta accttttata tttctctaca ggggcgcggc
      961 gtggggacaa ttcaacgcgt ctgtgagggg agcgtttccc tgctcgcagg tctgcagcga
     1021 ggagccgtaa tttttgcttc gcgccgtgcg gccatcaaaa tgtatggatg caaatgatta
     1081 tacatgggga tgtatgggct aaatgtacgg gcgacagtca catcatgccc ctgagctgcg
     1141 cacgtcaaga ctgtcaagga gggtattctg ggcctccatg tcgctggccg ggtgacccgg
     1201 cggggacgag gcaagctaaa cagatctggc gcgccttaat taaccctatg cggtgtgaaa
     1261 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     1321 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1381 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1441 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1501 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1561 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1621 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1681 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1741 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     1801 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     1861 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     1921 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc
     1981 gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgcc
     2041 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     2101 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2161 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2221 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3241 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3301 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3361 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3421 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3481 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3541 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3601 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3661 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3721 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3781 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3841 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3901 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3961 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4021 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4081 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4141 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4201 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4261 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4321 ctttcgtc
//
