LOCUS       sequence-49266-e        5539 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 5539)
  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 5539)
  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: pRSII319
FEATURES             Location/Qualifiers
     source          1..5539
                     /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(2001..2456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2138..2159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2350..2369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2582..2604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2596..2613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2597..2613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2620..2639
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2620..2638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2647..2754
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2671..2687
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2671..2687
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2721..2737)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2738)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2766..2784)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2805..2821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2805..2821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2818..2840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2829..2845
                     /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(2853..2859,2860..2877,2878..2883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2898..2919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3036..3053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3207..3795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3287..3306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3966..4757,4758..4826))
                     /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     4589..4608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4827..4931)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4968..5471
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5512..5530
                     /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 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgcga gtcggcgagc aaacagggtg ctgctactaa
      361 acgaacaata aagaaagaca atttgggcac ccaatataaa atgagcagtc aattcaatca
      421 attcagtgtg aaattttttt cttggaaaga tcattcactt ctacacagct gtacatgagt
      481 ttatatgaca gttttttttt atgtacatcc agttcaagat aatcagatta tatattggcc
      541 caaaagggat cattaacgaa ttgcgaaatg ggtaatctct ttacagttaa cagattctct
      601 ggtgattggc tttcctcaag gtagcaaata tactctaatt gtagcacgtc tactatgtat
      661 ttctttacct tgtcatagca cacacccact tgcgccacgc cgtgtgcgtc atcacttaga
      721 acaaatctgg agccacagtg cttcttgacc agattacata aggttttgct ggggtacggc
      781 tcctccaggc gctttcttaa tgcggacgta ttgatttcaa ttgcgccgcc ataggagtct
      841 ataaattgta aatttcttac aactgcatcg tatatttctg gccattcact gatgacgtcc
      901 agtgaagcta caggaactcc ggtttcttcg ttgcagttgc ccgatttctg gtttactagc
      961 atgtcattgg gcaaaaataa tttgtaaagg tcgaagtgac ccacgaccaa cggtttaata
     1021 ttgatcagca tttcgtactg tgattggaag taagacagga gaaaatgttt caaattatca
     1081 ttgaaggaat gcaatgaatt gtaccattgt tgttggtcga aatcaatagg tatcccgttg
     1141 acgtgatgga ccgaacccac acaaaacttc aaaatatcat tattctcctt catgagtcgc
     1201 tttgcatatt cgatatgagc catgtcacaa ctttcgatct ccattcctat aatgaattta
     1261 gtccgcacat cgggtctatc agcataacga gtcttgattt cttgcgcatg actcatgaaa
     1321 ttcttgaacg atgtttctag cttggttatg acttcctcag gattcttgcc caatgactgc
     1381 tcttcggggt atataaactt ggcctcaatt cttggtatgt gctctgtcaa acagtacgtg
     1441 tgaaagttga ggttgaccac ttgatcgacc acggaatcca aagggtccgt accgtgggca
     1501 ctatagtcac cggagtgtga atggtgtgag tgcatgacct tgaggatttt tactgaattg
     1561 tacgtatcaa gaagtactaa aatgtgtttg aaagggaaag attgtacggg aaatggaaaa
     1621 tgtacatata atatccagta tcatttgagt tgatagagat tatctctata gatagagaga
     1681 tttgacaaaa aaaataaaaa aatttaaatg gacgagtccg gaatcgaacc ggagacctct
     1741 cccatgctaa gggagcgcgc taccgactac gccacacgcc cgaacgtttg ttgaaatctt
     1801 ccaaacagaa atgatacagg tttactttac agcaagacat tagtagtttc attagcttat
     1861 caacatgtat gaaaccggca agattgaaag atgcattgta tactaaactc acaaattaga
     1921 gcttcaattt aattatatca gttattaccc tgcggtgtga aataccgcac agatgcgtaa
     1981 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2041 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2101 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2221 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     2461 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2521 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2581 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     2641 gaattggagc tccaccgcgg tggcggccgc tctagaacta gtggatcccc cgggctgcag
     2701 gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg taccagcttt
     2761 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     2821 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2881 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2941 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3001 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4561 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata
     4981 aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa
     5041 aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat
     5101 cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg
     5161 tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta
     5221 tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa
     5281 agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa
     5341 ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa
     5401 cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca
     5461 ggcaagcgat ccgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5521 atcacgaggc cctttcgtc
//
