LOCUS       sequence-287452-        6209 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 6209)
  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 6209)
  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..6209
                     /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..1393)
                     /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(2618..2650)
                     /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(2654..2671)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2681..2698)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2744..3028)
                     /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(2802..2821)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3055..3933
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             complement(join(5769..6209,1..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 gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagctta gagtcaacaa gctgaaaaca agtgaaacct
     1561 gagcaaccta gacgctagcg gcgacgggtg ctgcttggga ggtcaaacgc tcataggccg
     1621 cccgcatttt caggccggtc agcacttgga acagacccgt gccgttgttc gaacccgggt
     1681 aatctttgtg ctgcagcagc aggtgggtca actcaccgta gtactgcgtg tcggtcttgc
     1741 tgaggtggcc ctcgacgtag atcatctctt cgaggttctc gaactgaccg tcgacctcaa
     1801 gcactgagac ttcgtgaccg tagtaggtgt cagggccgta ggctaagcga atgcccggat
     1861 aggagcaggt cagcttacga ccgcagggcg tccactggat ggtcgaacct tcgtcgaaca
     1921 ggtaagccgg ctcaaaacca tcgcggcctt cccgttggat caactgcacc cgcagcacct
     1981 tgcgctcggt gtcattgggg attagctggg tcggcatgac gcggatgacg atgtccgcat
     2041 ggccacgctg gggctcaatg taggccttga agtcagggcg gcgcgcttcg atcgaggcga
     2101 ggacatcttc gtaggagtgg ccgcgttctg ccatatcgcg ttggatcttc caagcgattt
     2161 tgacttcgtc atcgatgtcg aggtaaacgc tgaaatcgag cagttcacgc acgcgctcgt
     2221 cgtaaagcgg atgcagaccc tcgatcacaa tgatgcgatt gggttcgatt ttttcgggcg
     2281 gatcgatcaa gccggtttca tggttgtaga tcggcttcat gatcgtttcg ccgttcttca
     2341 acgccttgac ctgttcatac atcaagtcaa agttgttggc gcggggatcc aaagccgtta
     2401 cgcctgcttc cttccggccc ttgcgatcga gactgtgata gtcatccaag cagatgaccg
     2461 tcatcaattc cgtaccaaac aagtcggcaa ggcgatttag gaaggttgat ttgccgcaac
     2521 cggagtcacc ggcaacgccg atcaaaacaa cacgatctgg cttgctcata ggatccctcg
     2581 agacagtgac gagctcggga tcgaattcgg atccgcgacc catttgctgt ccaccagtca
     2641 tgctagccat atggctgccg cgcggcacca ggccgctgct gtgatgatga tgatgatggc
     2701 tgctgcccat ggttaattcc tcctgttagc ccaaaaaacg ggtatggaga aacagtagag
     2761 agttgcgata aaaagcgtca ggtaggatcc gctaatctta tggataaaaa tgctatggca
     2821 tagcaaagtg tgacgccgtg caaataatca atgtggactt ttctgccgtg attatagaca
     2881 cttttgttac gcgtttttgt catggctttg gtcccgcttt gttacagaat gcttttaata
     2941 agcggggtta ccggtttggt tagcgagaag agccagtaaa agacgcagtg acggcaatgt
     3001 ctgatgcaat atggacaatt ggtttcttct ctgaatggcg ggagtatgaa aagtatggct
     3061 gaagcgcaaa atgatcccct gctgccggga tactcgttta atgcccatct ggtggcgggt
     3121 ttaacgccga ttgaggccaa cggttatctc gattttttta tcgaccgacc gctgggaatg
     3181 aaaggttata ttctcaatct caccattcgc ggtcaggggg tggtgaaaaa tcagggacga
     3241 gaatttgttt gccgaccggg tgatattttg ctgttcccgc caggagagat tcatcactac
     3301 ggtcgtcatc cggaggctcg cgaatggtat caccagtggg tttactttcg tccgcgcgcc
     3361 tactggcatg aatggcttaa ctggccgtca atatttgcca atacggggtt ctttcgcccg
     3421 gatgaagcgc accagccgca tttcagcgac ctgtttgggc aaatcattaa cgccgggcaa
     3481 ggggaagggc gctattcgga gctgctggcg ataaatctgc ttgagcaatt gttactgcgg
     3541 cgcatggaag cgattaacga gtcgctccat ccaccgatgg ataatcgggt acgcgaggct
     3601 tgtcagtaca tcagcgatca cctggcagac agcaattttg atatcgccag cgtcgcacag
     3661 catgtttgct tgtcgccgtc gcgtctgtca catcttttcc gccagcagtt agggattagc
     3721 gtcttaagct ggcgcgagga ccaacgtatc agccaggcga agctgctttt gagcaccacc
     3781 cggatgccta tcgccaccgt cggtcgcaat gttggttttg acgatcaact ctatttctcg
     3841 cgggtattta aaaaatgcac cggggccagc ccgagcgagt tccgtgccgg ttgtgaagaa
     3901 aaagtgaatg atgtagccgt caagttgtca taattggtaa cgaatcagac aattgacggc
     3961 ttgacggagt agcatagggt ttgcagaatc cctgcttcgt ccatttgaca ggcacattat
     4021 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     4081 cctaatgcag gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc
     4141 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     4201 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     4261 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     4321 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     4381 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     4441 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     4501 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     4561 gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta
     4621 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     4681 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     4741 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     4801 cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc
     4861 tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg
     4921 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     4981 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     5041 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     5101 tcccctacgt gctgctgaag ttgcccgcaa cagagagtgg aaccaaccgg tgataccacg
     5161 atactatgac tgagagtcaa cgccatgagc ggcctcattt cttattctga gttacaacag
     5221 tccgcaccgc tgccggtagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     5281 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg cccaacagtc
     5341 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgccctgc accattatgt
     5401 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     5461 cgaagcgcta accgttttta tcaggctctg ggaggcagaa taaatgatca tatcgtcaat
     5521 tattacctcc acggggagag cctgagcaaa ctggcctcag gcatttgaga agcacacggt
     5581 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
     5641 acgaccctgc cctgaaccga cgaccgggtc gaatttgctt tcgaatttct gccattcatc
     5701 cgcttattat cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct
     5761 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     5821 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     5881 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     5941 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     6001 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     6061 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     6121 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     6181 accagctcac cgtctttcat tgccatacg
//
