LOCUS       sequence-146159-        5127 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses A. thaliana PAL2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   McKenna R, Nielsen DR
  TITLE     Styrene biosynthesis from glucose by engineered E. coli.
  JOURNAL   Metab Eng. 2011 Sep;13(5):544-54. doi: 10.1016/j.ymben.2011.06.005. 
            Epub 2011 Jun 23.
  PUBMED    21722749
REFERENCE   2  (bases 1 to 5127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ymben.2011.06.005"; journalName: "Metab Eng"; date: 
            "2011-09"; volume: "13"; issue: "5"; pages: "544-54"
COMMENT     Sequence Label: pSpal2At
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1133..1154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1169..1174,1175..1192,1193..1199)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1207..1223
                     /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)."
     primer_bind     1212..1234
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1231..1247
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1231..1247
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3445..3462)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3445..3461)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3454..3476)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3736..4280
                     /direction=RIGHT
                     /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     4181..4200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4397..4414
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4806..4908
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4909..5127,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4975..4994)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
      541 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
      601 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
      661 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
      721 gagcctgata aaaacggtta gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgcttg tttcggcgtg
      841 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ctgccggcac ctgtcctacg
      901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
      961 cggaaggagc taccggacag cggtgcggac tgttgtaact cagaataaga aatgaggccg
     1021 ctcatggcgt tccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1081 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     1141 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1201 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat
     1261 tcaaggggat aaataatgga tcaaatcgaa gcaatgttgt gcggcggagg agagaagaca
     1321 aaagtggcgg ttactacgaa gactttggca gatccattga attggggttt agcagcggat
     1381 caaatgaaag gaagtcattt agatgaagtg aagaagatgg tcgaagagta tcgtagacca
     1441 gtcgtgaatc ttggcggaga aacactgacg atcggacaag ttgctgccat ctccaccgta
     1501 ggaggcagcg ttaaggttga gttagcggag acttcaagag ccggtgtgaa agctagcagt
     1561 gattgggtta tggagagcat gaacaaaggt actgacagtt acggagtcac caccggcttt
     1621 ggtgctactt ctcaccggag aaccaaaaac ggcaccgcat tacaaacaga actcattaga
     1681 tttttgaacg ccggaatatt cggaaacacg aaggagacat gtcacacact gccgcaatcc
     1741 gccacaagag ccgccatgct cgtcagagtc aacactcttc tccaaggata ctccgggatc
     1801 cgattcgaga tcctcgaagc gattacaagt ctcctcaacc acaacatctc tccgtcacta
     1861 cctctccgtg gaaccattac cgcctccggc gatctcgttc ctctctctta catcgccgga
     1921 cttctcaccg gccgtcctaa ttccaaagcc accggtcccg acggtgaatc gctaaccgcg
     1981 aaagaagctt ttgagaaagc cggaatcagt actggattct tcgatttaca acctaaggaa
     2041 ggtttagctc tcgttaatgg cacggcggtt ggatctggaa tggcgtcgat ggttctattc
     2101 gaagcgaatg tccaagcggt gttagcggag gttttatcag cgatcttcgc ggaggttatg
     2161 agcgggaaac ctgagtttac cgatcatctg actcatcgtt taaaacatca tcccggacaa
     2221 atcgaagcgg cggcgataat ggagcacata ctcgacggaa gctcatacat gaaattagct
     2281 caaaaggttc acgagatgga tccattgcag aaaccaaaac aagatcgtta cgctcttcgt
     2341 acatctcctc aatggctagg tcctcaaatt gaagtaatcc gtcaagctac gaaatcgata
     2401 gagcgtgaaa tcaactccgt taacgataat ccgttgatcg atgtttcgag gaacaaggcg
     2461 attcacggtg gtaacttcca aggaacacca atcggagttt ctatggataa cacgagattg
     2521 gcgattgctg cgattgggaa gctaatgttt gctcaattct ctgagcttgt taatgatttc
     2581 tacaacaatg gacttccttc gaatctaact gcttcgagta atccaagttt ggattatgga
     2641 ttcaaaggag cagagattgc tatggcttct tattgttctg agcttcaata cttggctaat
     2701 ccagtcacaa gccatgttca atcagctgag caacataatc aagatgtgaa ctctcttggt
     2761 ttgatctcgt ctcgtaaaac atctgaagct gtggatattc ttaagctaat gtcaacaacg
     2821 ttccttgtgg ggatatgtca agctgttgat ttgagacatt tggaggagaa tctgagacaa
     2881 actgtgaaga acacagtttc tcaagttgct aagaaagtgt taaccactgg aatcaacggt
     2941 gagttacatc cgtcaaggtt ttgcgagaag gacttgctta aggttgttga tcgtgagcaa
     3001 gtgttcacgt atgtggatga tccttgtagc gctacgtacc cgttgatgca gagactaaga
     3061 caagttattg ttgatcacgc tttgtccaac ggtgagactg agaagaatgc agtgacttcg
     3121 atctttcaaa agattggagc ttttgaagag gagcttaagg ctgtgcttcc aaaggaagtt
     3181 gaagcggcta gagcggctta tgggaatgga actgcgccga ttcctaaccg gattaaggaa
     3241 tgtaggtcgt atccgttgta taggttcgtg agggaagagc ttggaacgaa gttgttgact
     3301 ggagaaaagg ttgtgtctcc gggagaggag tttgataagg tcttcactgc tatgtgtgaa
     3361 ggtaaactta ttgatccgtt gatggattgt ctcaaggaat ggaacggagc tccgattccg
     3421 atttgctaag catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     3481 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3541 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3601 agcttatcga tgataagctg tcaaacatga gaattacaac ttatatcgta tggggctgac
     3661 ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt ttatctgatt
     3721 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
     3781 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg
     3841 aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc
     3901 ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg
     3961 cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4021 cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac
     4081 tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac
     4141 cgaaaggcag gaacaggaga gcgcacgagg gagccgccag gggaaacgcc tggtatcttt
     4201 atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag
     4261 gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat
     4321 cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag
     4381 tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat
     4441 attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca
     4501 ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgctga tgtccggcgg
     4561 tgcttttgcc gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa
     4621 cagaagccac tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca
     4681 tcacccgacg cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa
     4741 ataaatcctg gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga
     4801 gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     4861 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     4921 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     4981 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     5041 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     5101 cgcctgatga atgctcatcc ggaattt
//
