LOCUS       sequence-50973-e        5885 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 5885)
  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 5885)
  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: pRSII428
FEATURES             Location/Qualifiers
     source          1..5885
                     /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"
     rep_origin      complement(1511..1966)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1648..1669)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1860..1879
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1943..2323)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2092..2114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2106..2123
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2107..2123
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2133..2152
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2133..2151
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2160..2267
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2176..2192
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2177..2193
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2227..2243)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2227..2243)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2280..2300)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2280..2298)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2319..2335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2319..2335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2332..2354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2343..2359
                     /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(2367..2373,2374..2391,2392..2397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2412..2433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2550..2567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2721..3309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2801..2820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3480..4271,4272..4340))
                     /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     4103..4122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4341..4445)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4472..5818
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     5858..5876
                     /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 accatagatc tcttattcac gagtcagtct gactcttgcg agagatgagg atgtaataat
      241 actaatctcg aagatgccat ctaatacata tagacataca tatatatata tatacattct
      301 atatattctt acccagattc tttgaggtaa gacggttggg ttttatcttt tgcagttggt
      361 actattaaga acaatcgaat cataagcatt gcttacaaag aatacacata cgaaatatta
      421 acgataatgt caattacgaa gactgaactg gacggtatat tgccattggt ggccagaggt
      481 aaagttagag acatatatga ggtagacgct ggtacgttgc tgtttgttgc tacggatcgt
      541 atctctgcat atgacgttat tatggaaaac agcattcctg aaaagggtat cctattgacc
      601 aaactgtcag agttctggtt caagttcctg tccaacgatg ttcgtaatca tttggtagac
      661 atcgccccag gtaagactat tttcgattat ctacctgcaa aattgagcga accaaagtac
      721 aaaacgcaac tagaagaccg ctctctattg gttcacaaac ataaactaat tccattggaa
      781 gtaattgtca gaggctacat caccggatct gcttggaaag agtacgtaaa aacaggtact
      841 gtgcatggtt tgaaacaacc tcaaggactt aaagaatctc aagagttccc agaaccaatc
      901 ttcaccccat cgaccaaggc tgaacaaggt gaacatgacg aaaacatctc tcctgcccag
      961 gccgctgagc tggtgggtga agatttgtca cgtagagtgg cagaactggc tgtaaaactg
     1021 tactccaagt gcaaagatta tgctaaggag aagggcatca tcatcgcaga cactaaattc
     1081 gaatttggta ttgacgaaaa gaccaatgaa attattctag tggacgaggt gctaacgcca
     1141 gactcctcaa gattctggaa cggtgcctct tataaggtag gagaatccca agattcttac
     1201 gataagcaat ttttaagaga ctggcttact gctaataagt tgaacggtgt taacggcgtc
     1261 aaaatgcccc aagacattgt agacaggaca agggccaaat atatagaggc ttatgaaaca
     1321 ttgacagggt ctaaatggtc tcactaacgt gatttacata tactacaagt cgccagtgta
     1381 actcctcact gaatatgatt catacatacc cgtatgtatt aatgtataaa tgttctcaga
     1441 gcaaatttta tcgagatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     1501 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1561 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1621 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     1681 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     1741 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     1801 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     1861 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     1921 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     1981 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     2041 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     2101 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
     2161 gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc
     2221 ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     2281 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     2341 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2401 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2461 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2521 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3121 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4441 ccgcgcacat ttccccgaaa agtgccacct gaacgaagca tctgtgcttc attttgtaga
     4501 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4561 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     4621 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     4681 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     4741 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     4801 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     4861 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     4921 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     4981 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     5041 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     5101 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     5161 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     5221 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     5281 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     5341 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     5401 tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     5461 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     5521 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     5581 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     5641 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     5701 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     5761 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat
     5821 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5881 tcgtc
//
