LOCUS       sequence-50859-e        6861 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 6861)
  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 6861)
  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: pRSII322
FEATURES             Location/Qualifiers
     source          1..6861
                     /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"
     promoter        199..574
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     rep_origin      complement(2495..2950)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2632..2653)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2844..2863
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3076..3098
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3090..3107
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3091..3107
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3114..3133
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3114..3132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3141..3248
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3165..3181
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3165..3181
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3215..3231)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3216..3232)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3260..3278)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3299..3315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3299..3315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3312..3334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3323..3339
                     /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(3347..3353,3354..3371,3372..3377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3392..3413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3530..3547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3701..4289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3781..3800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4460..5251,5252..5320))
                     /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     5083..5102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5321..5425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5452..6798
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6838..6856
                     /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 tgaattaatt cttgaataat acataacttt tcttaaaaga atcaaagaca
      241 gataaaattt aagagatatt aaatattagt gagaagccga gaattttgta acaccaacat
      301 aacactgaca tctttaacaa cttttaatta tgatacattt cttacgtcat gattgattat
      361 tacagctatg ctgacaaatg actcttgttg catggctacg aaccgggtaa tactaagtga
      421 ttgactcttg ctgacctttt attaagaact aaatggacaa tattatggag catttcatgt
      481 ataaattggt gcgtaaaatc gttggatctc tcttctaagt acatcctact ataacaatca
      541 agaaaaacaa gaaaatcgga caaaacaatc aagtatggat tcaagaacag ttggtatatt
      601 aggaggggga caattgggac gtatgattgt tgaggcagca aacaggctca acattaagac
      661 ggtaatacta gatgctgaaa attctcctgc caaacaaata agcaactcca atgaccacgt
      721 taatggctcc ttttccaatc ctcttgatat cgaaaaacta gctgaaaaat gtgatgtgct
      781 aacgattgag attgagcatg ttgatgttcc tacactaaag aatcttcaag taaaacatcc
      841 caaattaaaa atttaccctt ctccagaaac aatcagattg atacaagaca aatatattca
      901 aaaagagcat ttaatcaaaa atggtatagc agttacccaa agtgttcctg tggaacaagc
      961 cagtgagacg tccctattga atgttggaag agatttgggt tttccattcg tcttgaagtc
     1021 gaggactttg gcatacgatg gaagaggtaa cttcgttgta aagaataagg aaatgattcc
     1081 ggaagcattg gaagtactga aggatcgtcc tttgtacgcc gaaaaatggg caccatttac
     1141 taaagaatta gcagtcatga ttgtgaggtc tgttaacggt ttagtgtttt cttacccaat
     1201 tgtagagact atccacaagg acaatatttg tgacttatgt tatgcgcctg ctagagttcc
     1261 ggactccgtt caacttaagg cgaagttgtt ggcagaaaat gcaatcaaat cttttcccgg
     1321 ttgtggtata tttggtgtgg aaatgttcta tttagaaaca ggggaattgc ttattaacga
     1381 aattgcccca aggcctcaca actctggaca ttataccatt gatgcttgcg tcacttctca
     1441 atttgaagct catttgagat caatattgga tttgccaatg ccaaagaatt tcacatcttt
     1501 ctccaccatt acaacgaacg ccattatgct aaatgttctt ggagacaaac atacaaaaga
     1561 taaagagcta gaaacttgcg aaagagcatt ggcgactcca ggttcctcag tgtacttata
     1621 tggaaaagag tcaagaccta acagaaaagt aggtcacata aatattattg cctccagtat
     1681 ggcggaatgt gaacaaaggc tgaactacat tacaggtaga actgatattc caatcaaaat
     1741 ctctgtcgct caaaagttgg acttggaagc aatggtcaaa ccattggttg gaatcatcat
     1801 gggatcagac tctgacttgc cggtaatgtc tgccgcatgt gcggttttaa aagattttgg
     1861 cgttccattt gaagtgacaa tagtctctgc tcatagaact ccacatagga tgtcagcata
     1921 tgctatttcc gcaagcaagc gtggaattaa aacaattatc gctggagctg gtggggctgc
     1981 tcacttgcca ggtatggtgg ctgcaatgac accacttcct gtcatcggtg tgcccgtaaa
     2041 aggttcttgt cttgatggag tagattcttt acattcaatt gtgcaaatgc ctagaggtgt
     2101 tccagtagct accgtcgcta ttaataatag tacgaacgct gcgctgttgg ctgtcagact
     2161 gcttggcgct tatgattcaa gttatacaac gaaaatggaa cagtttttat taaagcaaga
     2221 agaagaagtt cttgtcaaag cacaaaagtt agaaactgtc ggttacgaag catatcttga
     2281 aaacaagtaa tatataagtt tattgatata cttgtacagc aaataattat aaaatgatat
     2341 acctattttt taggctttgt tatgattaca tcaaatgtgg acttcataca tagaaatcaa
     2401 cgcttacagg tgtccttttt taagaatttc atacataaga tctatgcggt gtgaaatacc
     2461 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     2521 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2581 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2641 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2701 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2761 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2821 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2881 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2941 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3001 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3061 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
     3121 actcactata gggcgaattg gagctccacc gcggtggcgg ccgctctaga actagtggat
     3181 cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc
     3241 ccggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt
     3301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaacgaagca
     5461 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     5521 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     5581 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     5641 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     5701 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     5761 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     5821 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     5881 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     5941 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     6001 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     6061 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     6121 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     6181 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     6241 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     6301 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     6361 tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     6421 cctatacttt ctagctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa
     6481 aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc
     6541 acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt
     6601 ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt agtctagtac
     6661 ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag cactaccctt
     6721 tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca atgctatcat
     6781 ttcctttgat attggatcat ctaagaaacc attattatca tgacattaac ctataaaaat
     6841 aggcgtatca cgaggccctt t
//
