LOCUS       sequence-50877-e        4496 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 4496)
  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 4496)
  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: pRS40P
FEATURES             Location/Qualifiers
     source          1..4496
                     /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..1350)
                     /product="yeast selectable marker conferring bialaphos 
                     resistance (Goldstein and McCusker, 1999)"
                     /label=patMX4
     terminator      complement(249..446)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(500..952)
                     /codon_start=1
                     /gene="Streptomyces viridochromogenes pat1"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialaphos or phosphinothricin"
                     /translation="MAAVCDIVNHYIETSTVNFRTEPQTPQEWIDDLERLQDRYPWLVA
                     EVEGVVAGIAYAGPWKARNAYDWTVESTVYVSHRHQRLGLGSNLYTHLLKSMEAQGFKS
                     VVAVIGLPNDPSVRLHEALGYTARGTLRAAGYKHGGWHDVGFWQRDF"
     promoter        complement(1007..1350)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     rep_origin      complement(1467..1922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1604..1625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1816..1835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1899..2279)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2048..2070
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2062..2079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2063..2079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2089..2108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2089..2107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2116..2223
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2132..2148
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2133..2149
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2183..2199)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2183..2199)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2236..2256)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2236..2254)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2275..2291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2275..2291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2288..2310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2299..2315
                     /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(2323..2329,2330..2347,2348..2353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2368..2389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2506..2523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2677..3265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2757..2776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3436..4227,4228..4296))
                     /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     4059..4078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4297..4401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4469..4487
                     /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 taccttctgc
      301 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
      361 acatttgatt agaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
      421 aaaacaagaa tctttttatt gtcagtactg attagatctg tgtgacgggc cggacggggc
      481 ggagcggggc cggcagctcg aagtcgcgct gccagaaccc cacgtcgtgc cagcccccgt
      541 gcttgtagcc ggctgcccgc agcgtcccgc gcgcggtgta tccgagcgcc tcgtgcaggc
      601 gcacgctcgg gtcgttgggc agtccgatga cggcgaccac gctcttgaag ccctgggcct
      661 ccatggactt cagcaggtgg gtgtagaggt tggagcccag tccgagccgc tggtgccggt
      721 gggagacgta caccgtcgac tcgacggtcc agtcgtaggc gttgcgggcc ttccaggggc
      781 cggcgtaggc gatgccggcg acgacgccct ccacctcggc gacgagccag gggtagcggt
      841 cctggaggcg ctccaggtcg tcgatccact cctgcggagt ctgcggctcc gtacggaagt
      901 tgaccgtgct cgtctcgatg tagtgattga cgatgtcgca gaccgccgcc atgtcggcgg
      961 cggtggcggg acggatctcg accgggcgtc gttctgggct acccatggtt gtttatgttc
     1021 ggatgtgatg tgagaactgt atcctagcaa gattttaaaa ggaagtatat gaaagaagaa
     1081 cctcagtggc aaatcctaac cttttatatt tctctacagg ggcgcggcgt ggggacaatt
     1141 caacgcgtct gtgaggggag cgtttccctg ctcgcaggtc tgcagcgagg agccgtaatt
     1201 tttgcttcgc gccgtgcggc catcaaaatg tatggatgca aatgattata catggggatg
     1261 tatgggctaa atgtacgggc gacagtcaca tcatgcccct gagctgcgca cgtcaagact
     1321 gtcaaggagg gtattctggg cctccatgtc gctggccggg tgacccggcg gggacgaggc
     1381 aagctaaaca gatctggcgc gccttaatta accctatgcg gtgtgaaata ccgcacagat
     1441 gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc
     1501 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1561 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1621 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1681 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1741 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     1801 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     1861 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     1921 gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1981 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     2041 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     2101 ctatagggcg aattgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat
     2161 atcgaattcc tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag
     2221 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2341 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4441 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
