LOCUS       sequence-50974-e        6370 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 6370)
  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 6370)
  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: pRSII329
FEATURES             Location/Qualifiers
     source          1..6370
                     /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(2000..2455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2137..2158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2349..2368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2581..2603
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2595..2612
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2596..2612
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2619..2638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2619..2637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2646..2753
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2670..2686
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2670..2686
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2720..2736)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2721..2737)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2765..2783)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2804..2820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2804..2820)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2817..2839)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2828..2844
                     /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(2852..2858,2859..2876,2877..2882))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2897..2918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3035..3052)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3206..3794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3286..3305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3965..4756,4757..4825))
                     /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     4588..4607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4826..4930)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4957..6303
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6343..6361
                     /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 atgcatgtat actaaactca caaattagag
     1921 cttcaattta attatatcag ttattaccct gcggtgtgaa ataccgcaca gatgcgtaag
     1981 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2041 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2101 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2161 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2221 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2281 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2341 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     2401 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     2461 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     2521 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     2581 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     2641 aattggagct ccaccgcggt ggcggccgct ctagaactag tggatccccc gggctgcagg
     2701 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt accagctttt
     2761 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
     2821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3001 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3061 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4921 gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt
     4981 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     5041 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     5101 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     5161 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     5221 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     5281 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     5341 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     5401 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     5461 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     5521 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     5581 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     5641 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     5701 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     5761 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     5821 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     5881 gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     5941 tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga gcgcttccga
     6001 aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta tatctgcgtg
     6061 ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg cttaaatgcg
     6121 tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct gtgatattat
     6181 cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct gttctatatg
     6241 ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct ttgatattgg
     6301 atcatctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6361 ccctttcgtc
//
