LOCUS       sequence-228913-        6390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  URA3-marked RPS28.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6390)
  AUTHORS   McGeachy AM, Meacham ZA, Ingolia NT
  TITLE     An Accessible Continuous-Culture Turbidostat for Pooled Analysis of 
            Complex Libraries.
  JOURNAL   ACS Synth Biol. 2019 Apr 19;8(4):844-856. doi: 
            10.1021/acssynbio.8b00529. Epub 2019 Apr 2.
  PUBMED    30908907
REFERENCE   2  (bases 1 to 6390)
  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"; doi: 
            "10.1021/acssynbio.8b00529"; journalName: "ACS Synth Biol"; date: 
            "2019-04-19- 19"; volume: "8"; issue: "4"; pages: "844-856"
COMMENT     Sequence Label: pRPL28 CaURA3 (pNTI592)
FEATURES             Location/Qualifiers
     source          1..6390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..80)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(62..80)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(167..184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(338..926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(418..437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1097..1888,1889..1957))
                     /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     1720..1739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1958..2062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2130..2148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2186..2208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2308..2327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2408..2426
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2408..2425
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    2470..2973
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     terminator      4723..4910
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4778..4802)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        4985..5328
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      6147..6344
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct
       61 ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc
      121 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
      181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1021 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1981 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2041 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2101 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     2161 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2221 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2281 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2341 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     2401 acatacgatt taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag
     2461 gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat
     2521 aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag
     2581 atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct
     2641 ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc
     2701 ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct
     2761 tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta
     2821 tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat
     2881 acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga
     2941 tacgaggcgc gtgtaagtta caggcaagcg atccgtccgg atccaaatta tgccaccgtt
     3001 accttacgtt tcatggttaa tcatcgttta ctgccgccta tgagcgtaag ctaatgttat
     3061 aaagaaacaa gctataatat tgttaaatat agttgatcaa cagcattgta atgattacaa
     3121 gagacgaggt ggaatgaacc ttatgaaatg cgtattatat ataaactgta ataagagcta
     3181 agttgaattg aaatctacga tacttgatgt tgacattata gcactagttc ccaggaaacc
     3241 ctttcgaaaa acacagcaaa aacaagagta ctgtaaccaa tgtaacatct gtacaccagg
     3301 gacccacaca ttaccaaaat caaaattatt tttctaatgc ctgttatttt tcctattttt
     3361 cctctggcgc gtgaatagcc cgcagagacg caaacaattt tcctcgcagt ttttcgcttg
     3421 tttaatgcgt attttcccag ataggttcaa acctttcatc tgtatcccgt atatttaaga
     3481 tggcgtttgc tttctccgtt gatttttttc cttcttagtg atttttttgc attaaatccc
     3541 agaacaatca tccaactaat caagaatgcc ttccagattc actaagacta gaaagcacag
     3601 aggtcacgtc tcaggtatgt agttccattt ggaagaggga atgaaagaac caagacggtg
     3661 actttttttt tagtgttgtg caaccaatat gtcgtgtgta tatcatggta caggagaatg
     3721 tcaatcagct aagtgtactc aacatatttc tttgtgtttt gattgcgaac tttgtattac
     3781 catctcactg ttgagacggc ttatttgagg taatagctcg agtaaatgta ctcttccatc
     3841 gcaaactgag caaaaagaaa gtgtgcatag cctttgtcat acttctcctt tattatacca
     3901 tgatattcag aacagtcata ctgtctactc attttacggc tataaaaggt aactttcatt
     3961 tagattatgg aaagcactaa ttatcgctgt atggtaccaa atggttgtag agagcgcaat
     4021 tatgaaaaag agttaccacg tttcttttgt ttcgataaaa tgtccagttg aaaacctgtt
     4081 ttactaacga tttaaaaatt gtatttcatt acaatatttt ttttgtacag ccggtaaagg
     4141 tcgtatcggt aagcacagaa agcaccccgg tggtagaggt atggccggtg gtcaacatca
     4201 ccacagaatt aacatggata aataccatcc aggttatttc ggtaaggttg gtatgagata
     4261 cttccacaag caacaagctc atttctggaa gccagtcttg aacttggaca aattgtggac
     4321 attgatccca gaagacaaga gagaccaata cttgaaatct gcttctaagg aaactgctcc
     4381 agttattgac actttggcag ccggttacgg taagatcttg ggtaagggta gaattccaaa
     4441 tgttccagtt atcgtcaaag ctagattcgt ctccaagttg gctgaagaaa aaatcagagc
     4501 tgctggtggt gttgttgaat tgatcgctta agcgcatcaa caaaaactct atgtattttc
     4561 caataaatta tatatcttca gtttaatcta attcaacatc tacttctgta ttatttctat
     4621 cacccatttt caccgttttt tcgtcttgtc tattttgaag ttaccagtgg cagatcccaa
     4681 cgcttttatc gcttttttaa aaggcgcgcc acttctaaat aagcgaattt cttatgattt
     4741 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     4801 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     4861 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
     4921 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     4981 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     5041 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     5101 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcggacctg
     5161 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     5221 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     5281 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gacagtcaac
     5341 actaagacct atagtgagag agcagaaact catgcctcac cagtagcaca acgattattt
     5401 cgattaatgg aactgaagaa aaccaattta tgtgcatcaa ttgatgttga taccactaag
     5461 gaattccttg aattaattga taaattgggt ccttatgtat gcttaatcaa gactcatatt
     5521 gatataatca atgatttttc ctatgaatcc actattgaac cattattaga actttcacgt
     5581 aaacatcaat ttatgatttt tgaagataga aaatttgctg atattggtaa taccgtgaag
     5641 aaacaatata ttggtggagt ttataaaatt agtagttggg cagatattac taatgctcat
     5701 ggtgtcactg ggaatggagt agttgaagga ttaaaacagg gagctaaaga aaccaccacc
     5761 aaccaagagc caagagggtt attgatgtta gctgaattat catcagtggg atcattagca
     5821 tatggagaat attctcaaaa aactgttgaa attgctaaat ccgataagga atttgttatt
     5881 ggatttattg cccaacgtga tatgggtgga caagaagaag gatttgattg gcttattatg
     5941 acacctggag ttggattaga tgataaaggt gatggattag gacaacaata tagaactgtt
     6001 gatgaagttg ttagcactgg aactgatatt atcattgttg gtagaggatt gtttggtaaa
     6061 ggaagagatc cagatattga aggtaaaagg tatagagatg ctggttggaa tgcttatttg
     6121 aaaaagactg gccaattata atcagtactg acaataaaaa gattcttgtt ttcaagaact
     6181 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     6241 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     6301 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     6361 aacgccgcca tccagtgtcg aaaacgagct
//
