LOCUS       sequence-43388-e        4960 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 4960)
  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 4960)
  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: pRS40H
FEATURES             Location/Qualifiers
     source          1..4960
                     /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"
     gene            complement(249..1824)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(249..446)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(452..1480)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(452..1480)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(1481..1824)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     rep_origin      complement(1941..2396)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2078..2099)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2290..2309
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2373..2753)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2522..2544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2536..2553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2537..2553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2563..2582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2563..2581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2590..2697
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2606..2622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2607..2623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2657..2673)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2657..2673)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2710..2730)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2710..2728)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2749..2765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2749..2765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2762..2784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2773..2789
                     /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(2797..2803,2804..2821,2822..2827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2842..2863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2980..2997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3151..3739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3231..3250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3910..4701,4702..4770))
                     /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     4533..4552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4771..4875)
                     /gene="bla"
                     /label=AmpR promoter
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 attattcctt tgccctcgga cgagtgctgg
      481 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc
      541 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
      601 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
      661 ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat
      721 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
      781 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
      841 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
      901 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
      961 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
     1021 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
     1081 acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct
     1141 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
     1201 gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg
     1261 ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
     1321 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
     1381 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     1441 tctcgacaga cgtcgcggtg agttcaggct ttttacccat ggttgtttat gttcggatgt
     1501 gatgtgagaa ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag
     1561 tggcaaatcc taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc
     1621 gtctgtgagg ggagcgtttc cctgctcgca ggtccgcagc gaggagccgt aatttttgct
     1681 tcgcgccgtg cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg
     1741 ctaaatgtac gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag
     1801 gagggtattc tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta
     1861 aacagatctg gcgcgcctta attaacccta tgcggtgtga aataccgcac agatgcgtaa
     1921 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1981 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2041 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2101 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2161 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2221 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2281 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2341 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     2401 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2461 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2521 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     2581 ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgatatcgaa
     2641 ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt ggagctccag
     2701 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     2761 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2821 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2881 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2941 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3001 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3061 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3901 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4201 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4741 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4921 tatcatgaca ttaacctata aaaataggcg tatcacgagg
//
