LOCUS       sequence-54361-e        6211 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 6211)
  AUTHORS   Parikh MR, Greene DN, Woods KK, Matsumura I
  TITLE     Directed evolution of RuBisCO hypermorphs through genetic selection 
            in engineered E.coli.
  JOURNAL   Protein Eng Des Sel. 2006 Mar;19(3):113-9. Epub 2006 Jan 19.
  PUBMED    16423843
REFERENCE   2  (bases 1 to 6211)
  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: "Protein Eng
            Des Sel."; date: "2006-03"; volume: "19(3)"; pages: "113-9. Epub 
            2006 Jan 19"
COMMENT     Sequence Label: PBAD-his6-prkA-pACYC184
FEATURES             Location/Qualifiers
     source          1..6211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1392)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2620..2652)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(2656..2673)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2683..2700)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2746..3030)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(2804..2823)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3057..3935
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             complement(join(5771..6211,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct
      961 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1021 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1081 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1141 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1201 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1261 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1321 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1381 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1441 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     1501 tgtttgacag cttatcatcg ataagcttag agtcaacaag ctgaaaacaa gtgaaacctg
     1561 agcaacctag acgctagcgg cgacgggtgc tgcttgggag gtcaaacgct cataggccgc
     1621 ccgcattttc aggccggtca gcacttggaa cagacccgtg ccgttgttcg aacccgggta
     1681 atctttgtgc tgcggcagca ggtgggtcaa ctcaccgtag tactgcgtgt cggtcttgct
     1741 gaggtggccc tcgacgtaga tcatctcttc gaggttctcg aactgaccgt cgacctcaag
     1801 cactgagact ccgtgaccgt agtaggtgcc agggccgtag gctaagcgaa tgcccggata
     1861 ggagcaggtc agcttacgac cgcagggcgt ccactggatg gtcgaacctt cgtcgaacag
     1921 gtaagccggc tcaaaaccat cgcggccttc ccgttggatc aactgcaccc gcagcacctt
     1981 gcgctcggtg tcattgggga ttagctgggt cggcatgacg cggatgacga tgtccgcatg
     2041 gccacgctgg ggctcaatgt aggccttgaa gtcagggcgg cgcgcttcga tcgaggcgag
     2101 gacatcttcg taggagtggc cgcgttctgc catatcgcgt tggatcttcc aagcgatttt
     2161 gacttcgtca tcgatgtcga ggtaaacgct gaaatcgagc agttcacgca cgcgctcgtc
     2221 gtaaagcgga tgcagaccct cgatcacaat gatgcgattg ggttcgattt tttcgggcgg
     2281 atcgatcaag ccggtttcat ggttgtagat cggcttcatg atcgtttcgc cgttcttcaa
     2341 cgccttgacc tgttcataca tcaagtcaaa gttgttggcg cggggatcca aagccgttac
     2401 gcctgcttcc ttccggccct tgcgatcgag actgtgatag tcatccaagc agatgaccgt
     2461 catcaattcc gtaccaaaca agtcggcaag gcgatttagg aaggttgatt tgccgcaacc
     2521 ggagtcaccg gcaacgccga tcagaacaac acgatctggc ttgctcatag gatccctcga
     2581 gacagtgacg agctcgggcc catcgaattc ggatccgcga cccatttgct gtccaccagt
     2641 catgctagcc atatggctgc cgcgcggcac caggccgctg ctgtgatgat gatgatgatg
     2701 gctgctgccc atggttaatt cctcctgtta gcccaaaaaa cgggtatgga gaaacagtag
     2761 agagttgcga taaaaagcgt caggtaggat ccgctaatct tatggataaa aatgctatgg
     2821 catagcaaag tgtgacgccg tgcaaataat caatgtggac ttttctgccg tgattataga
     2881 cacttttgtt acgcgttttt gtcatggctt tggtcccgct ttgttacaga atgcttttaa
     2941 taagcggggt taccggtttg gttagcgaga agagccagta aaagacgcag tgacggcaat
     3001 gtctgatgca atatggacaa ttggtttctt ctctgaatgg cgggagtatg aaaagtatgg
     3061 ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt taatgcccat ctggtggcgg
     3121 gtttaacgcc gattgaggcc aacggttatc tcgatttttt tatcgaccga ccgctgggaa
     3181 tgaaaggtta tattctcaat ctcaccattc gcggtcaggg ggtggtgaaa aatcagggac
     3241 gagaatttgt ttgccgaccg ggtgatattt tgctgttccc gccaggagag attcatcact
     3301 acggtcgtca tccggaggct cgcgaatggt atcaccagtg ggtttacttt cgtccgcgcg
     3361 cctactggca tgaatggctt aactggccgt caatatttgc caatacgggg ttctttcgcc
     3421 cggatgaagc gcaccagccg catttcagcg acctgtttgg gcaaatcatt aacgccgggc
     3481 aaggggaagg gcgctattcg gagctgctgg cgataaatct gcttgagcaa ttgttactgc
     3541 ggcgcatgga agcgattaac gagtcgctcc atccaccgat ggataatcgg gtacgcgagg
     3601 cttgtcagta catcagcgat cacctggcag acagcaattt tgatatcgcc agcgtcgcac
     3661 agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt ccgccagcag ttagggatta
     3721 gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc gaagctgctt ttgagcacca
     3781 cccggatgcc tatcgccacc gtcggtcgca atgttggttt tgacgatcaa ctctatttct
     3841 cgcgggtatt taaaaaatgc accggggcca gcccgagcga gttccgtgcc ggttgtgaag
     3901 aaaaagtgaa tgatgtagcc gtcaagttgt cataattggt aacgaatcag acaattgacg
     3961 gcttgacgga gtagcatagg gtttgcagaa tccctgcttc gtccatttga caggcacatt
     4021 atgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     4081 ttcctaatgc aggagtcgca taagggagag cgtcgaccga tgcccttgag agccttcaac
     4141 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     4201 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     4261 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     4321 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     4381 gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg
     4441 cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc
     4501 gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga
     4561 tcgctcgcgg ctcttaccag cctaacttcg atcactggac cgctgatcgt cacggcgatt
     4621 tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac
     4681 cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg
     4741 gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct
     4801 tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca
     4861 tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca
     4921 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     4981 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     5041 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     5101 agtcccctac gtgctgctga agttgcccgc aacagagagt ggaaccaacc ggtgatacca
     5161 cgatactatg actgagagtc aacgccatga gcggcctcat ttcttattct gagttacaac
     5221 agtccgcacc gctgtccggt agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     5281 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgcccaaca
     5341 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgccc tgcaccatta
     5401 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     5461 taacgaagcg ctaaccgttt ttatcaggct ctgggaggca gaataaatga tcatatcgtc
     5521 aattattacc tccacgggga gagcctgagc aaactggcct caggcatttg agaagcacac
     5581 ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat
     5641 ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc
     5701 atccgcttat tatcacttat tcaggcgtag caccaggcgt ttaagggcac caataactgc
     5761 cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     5821 ttctgccgac atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca
     5881 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     5941 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     6001 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     6061 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     6121 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     6181 tcaccagctc accgtctttc attgccatac g
//
