LOCUS       sequence-50581-e        5049 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5049)
  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 5049)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRSII318
FEATURES             Location/Qualifiers
     source          1..5049
                     /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"
     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     2130..2149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2130..2148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2157..2264
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2181..2197
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2181..2197
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2231..2247)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2232..2248)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2276..2294)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2315..2331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2315..2331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2328..2350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2339..2355
                     /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(2363..2369,2370..2387,2388..2393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2408..2429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2546..2563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2717..3305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2797..2816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3476..4267,4268..4336))
                     /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     4099..4118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4337..4441)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4478..4981
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5022..5040
                     /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 agtgaattgt aatacgactc actatagggc gaattggagc
     2161 tccaccgcgg tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattcgata
     2221 tcaagcttat cgataccgtc gacctcgagg gggggcccgg taccagcttt tgttcccttt
     2281 agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2341 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2401 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2461 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2521 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2581 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2641 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2881 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3121 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4441 gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta
     4501 taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta
     4561 aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa
     4621 tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg
     4681 tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg
     4741 aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt
     4801 ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc
     4861 ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa
     4921 attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat
     4981 ccgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5041 cctttcgtc
//
