LOCUS       sequence-83691-e        8353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains crtE, crtB, and idi genes of Erwinia herbicola (Pantoea 
            agglomerans) Eho10, and pds (crtP) gene of Synechococcus PCC7942, 
            and thereby produces zeta-carotene in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8353)
  AUTHORS   Cunningham FX Jr, Gantt E
  TITLE     A portfolio of plasmids for identification and analysis of 
            carotenoid pathway enzymes: Adonis aestivalis as a case study.
  JOURNAL   Photosynth Res. 2007 May;92(2):245-59. Epub 2007 Jul 17.
  PUBMED    17634749
REFERENCE   2  (bases 1 to 8353)
  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: "Photosynth 
            Res."; date: "2007-05"; volume: "92(2)"; pages: "245-59. Epub 2007 
            Jul 17"
COMMENT     Sequence Label: pAC-ZETAipi
FEATURES             Location/Qualifiers
     source          1..8353
                     /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"
     promoter        join(1505..1510,1511..1527,1528..1533)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     protein_bind    4124..4145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4160..4165,4166..4183,4184..4190)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4198..4214
                     /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     4203..4225
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4222..4238
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4222..4238
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4257..4277
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4259..4277
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      5425..5434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(7913..8353,1..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 gataagcttt aatgcggtag tttatcacag ttaaattgct
     1561 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg
     1621 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg
     1681 atatcgtcca ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt
     1741 tgatgcaatt tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc
     1801 cagtcctgct cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac
     1861 ccgtcctgtg gatctaaagg cacagcgtct catgcttcgc acaatgtaaa actgcttcag
     1921 aacctggcga gagctatccg cgcggtctac ggttaactga tactaaaaga caattcagcg
     1981 ggtaaccttg caatggtgag tggcagtaaa gcgggcgttt cgcctcatcg cgaaatagaa
     2041 gtaatgagac aatccattga cgatcacctg gctggcctgt tacctgaaac cgacagccag
     2101 gatatcgtca gccttgcgat gcgtgaaggc gtcatggcac ccggtaaacg gatccgtccg
     2161 ctgctgatgc tgctggccgc ccgcgacctc cgctaccagg gcagtatgcc tacgctgctc
     2221 gatctcgcct gcgccgttga actgacccat accgcgtcgc tgatgctcga cgacatgccc
     2281 tgcatggaca acgccgagct gcgccgcggt cagcccacta cccacaaaaa atttggtgag
     2341 agcgtggcga tccttgcctc cgttgggctg ctctctaaag cctttggtct gatcgccgcc
     2401 accggcgatc tgccggggga gaggcgtgcc caggcggtca acgagctctc taccgccgtg
     2461 ggcgtgcagg gcctggtact ggggcagttt cgcgatctta acgatgccgc cctcgaccgt
     2521 acccctgacg ctatcctcag caccaaccac ctcaagaccg gcattctgtt cagcgcgatg
     2581 ctgcagatcg tcgccattgc ttccgcctcg tcgccgagca cgcgagagac gctgcacgcc
     2641 ttcgccctcg acttcggcca ggcgtttcaa ctgctggacg atctgcgtga cgatcacccg
     2701 gaaaccggta aagatcgcaa taaggacgcg ggaaaatcga cgctggtcaa ccggctgggc
     2761 gcagacgcgg cccggcaaaa gctgcgcgag catattgatt ccgccgacaa acacctcact
     2821 tttgcctgtc cgcagggcgg cgccatccga cagtttatgc atctgtggtt tggccatcac
     2881 cttgccgact ggtcaccggt catgaaaatc gcctgatacc gcccttttgg gttcaagcag
     2941 tacataacga tggaaccaca ttacaggagt agtgatgaat gaaggacgag cgccttgttc
     3001 agcgtaagaa cgatcatctg gatatcgttc tcgacccccg tcgcgccgta actcaggcta
     3061 gcgcaggttt tgagcgctgg cgctttaccc actgcgccct gccagagctg aattttagcg
     3121 acatcacgct ggaaaccacc ttcctgaatc ggcagctaca ggctccgctg ctgatcagct
     3181 ccatgaccgg cggcgttgag cgctcgcgcc atatcaaccg ccacctcgcc gaggcggcgc
     3241 aggtgctaaa aattgcgatg ggggtgggct cccagcgcgt cgccattgag agcgacgcgg
     3301 gcttagggct ggataaaacc ctgcggcagc tggctccgga cgtgccgctg ctggcgaacc
     3361 tcggcgcggc gcagctgacc ggcagaaaag gtattgatta cgcccgacgg gccgtggaga
     3421 tgatcgaggc ggatgcgctg attgtgcacc taaacccgct gcaggaggcg ctacagcccg
     3481 gcggcgatcg cgactggcgc ggacggctgg cggctattga aactctggtc cgcgagctgc
     3541 ccgttccgct ggtggtgaaa gaggtgggag ccggtatctc ccgaaccgtg gccgggcagc
     3601 tgatcgatgc cggcgttacc gtgattgacg tcgcgggcgc gggcggcacc agctgggccg
     3661 ccgttgaagg cgagcgggcg gccaccgagc agcagcgcag cgtggccaac gtctttgccg
     3721 actgggggat ccccaccgct gaggcgctgg ttgacattgc cgaggcctgg ccgcagatgc
     3781 cccttattgc ctcgggcggg attaaaaacg gcgtcgacgc ggcgaaagcg ctgcggctcg
     3841 gcgcgtgcat ggtagggcag gccgccgccg tgctcggcag cgcaggcgtc tccacggaga
     3901 aggtgatcga tcacttcaac gtgattattg agcagctgcg ggtggcctgc ttctgcaccg
     3961 gcagccgcag cctgagcgat ctaaagcagg ctgatatccg ctatgtgcgg gatacgccat
     4021 gagccatttt gccattgtgg caccgccgct ctacagtcat gcggtggcgc tgctggcacg
     4081 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     4141 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     4201 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     4261 ttaaccctca ctaaagggaa caaaagctgg attcttctgc ccttccagcc aaaagccatg
     4321 cgcgtagcga tcgccggtgc cggacttgcc ggactctcct gtgccaagta cttggccgat
     4381 gccggtcata cgcccatcgt ctatgaacgt cgggacgtcc ttggcggcaa ggttgccgct
     4441 tggaaagatg aagacggcga ctggtacgaa actggcctac atatcttttt tggggcttac
     4501 cccaacatgt tgcagctctt taaggagctg aacattgaag atcgcctgca gtggaagtcc
     4561 cactcgatga tcttcaacca acccacaaag ccgggcacct attcgcgctt cgacttccca
     4621 gacattccag cgccaatcaa cggtgttgca gcaatcctca gcaacaacga catgttgacc
     4681 tgggaagaaa aaatcaagtt tggcttgggc ttgttgccag cgatgattcg cggccagtcc
     4741 tacgtcgaag agatggatca atactcatgg acggagtggc tgcgcaaaca aaatattcca
     4801 gagcgggtca acgatgaagt cttcatcgcc atggctaaag cgctcaactt tattgacccg
     4861 gacgaaattt ccgccacggt cgtcctaacg gcactcaacc gcttcttgca agagaagaaa
     4921 ggttcaatga tggccttttt ggatggtgcg ccgcccgagc gtctttgcca gccgatcgtc
     4981 gaacatgtcc aagctcgcgg tggtgatgtg ctgctgaatg cgcctctgaa agagttcgtg
     5041 ctcaatgacg acagtagcgt ccaagctttt cggattgctg gcatcaaagg tcaagaagaa
     5101 caactcattg aggcagatgc ctacgtttcg gcactgccgg ttgatccgct caagctactg
     5161 ttgccggatg catggaaagc catgccctac ttccagcaac tcgatggtct gcagggcgtg
     5221 ccggtcatca acattcacct ctggttcgat cgcaagctga ccgatatcga tcacctgctg
     5281 ttctcgcgat cgcccctgct cagtgtctat gccgacatga gtaacacctg tcgcgagtac
     5341 gaagatcccg atcgctcaat gctagagctg gtcttcgccc ccgccaaaga ctggattggc
     5401 cgctccgacg aagacatctt ggctgccacc atggccgaga ttgaaaagct attcccacag
     5461 catttcagcg gtgagaatcc ggcacgtctg cgcaaataca aaattgtcaa aacgcccctg
     5521 tcggtctaca aagccacgcc gggccgtcaa caatatcgcc ccgatcaagc tagcccgatc
     5581 gctaatttct tcctgaccgg cgactacacc atgcagcgct acctcgccag tatggaaggg
     5641 gcggtcctat ctggtaagct gacagcgcaa gccatcattg ctcgccaaga tgagttgcaa
     5701 cgtcgcagca gcggacgacc gctggccgcg agtcaggcat agacactgtt acccatgttg
     5761 cagttgctag ctctgaagct tcactggtca ccctctcacc cccgactcgc accgtctgtc
     5821 tgatgctgca gatgataccg gcacggcctt cacgggcgct cgcctctctg tcggaggcct
     5881 acgaggagtg ccgacaaatt accgctcgct acgctaagac cttttacctc gggacgcttt
     5941 tgatgcccga ggccaagcgt caggccatct gggccatcta cgtctggtgt cgtcgtaccg
     6001 atgaactagt cgatggccct caagcagcgc agactaactt cgcgaccctg gatgcttggg
     6061 agcggcggtt ggaacgactg tttgctgggg aacccgaaga cgactgcgat gttgccttgg
     6121 tggatacctt ggctcgctat cctctcgata tccagccctt ccgcgacatg atcgaagggc
     6181 agcggatgga tctgctgcaa aaccgctatt ccacttttga ggatctctac acctactgct
     6241 accgtgtcgc cggtactgtg ggactgatgt cacagccggt gatgggcatt gaatcgacca
     6301 atagtcgtgc cccttgggat ctttaccggc agcgcgctgg cggatgactt ctcgctctac
     6361 ctgcactcgc cctgcgtgac cgatccctcg ctcgcgcctc ccggctgcgc cagcttctac
     6421 gtgctggccc cggtgccgca tcttggcaac gcgccgctgg actgggcgca ggaggggccg
     6481 aagctgcgcg accgcatctt tgactacctt gaagagcgct atatgcccgg cctgcgtagc
     6541 cagctggtga cccagcggat ctttaccccg gcagacttcc acgacacgct ggatgcgcat
     6601 ctgggatcgg ccttctccat cgagccgctg ctgacccaaa gcgcctggtt ccgcccgcac
     6661 aaccgcgaca gcgacattgc caacctctac ctggtgggcg caggtactca ccctggggcg
     6721 ggcattcctg gcgtagtggc ctcggcgaaa gccaccgcca gcctgatgat tgaggatctg
     6781 caatgagcca accgccgctg cttgaccacg ccacgcagac catggccaac ggctcgaaaa
     6841 gttttgccac cgctgcgaag ctgttcgacc cggccacccg ccgtagcgtg ctgatgctct
     6901 acacctggtg ccgccactgc gatgacgtca ttgacgacca gacccacggc ttcgccagcg
     6961 aggccgcggc ggaggaggag gccacccagc gcctggcccg gctgcgcacg ctgaccctgg
     7021 cggcgtttga aggggccgag atgcaggatc cggccttcgc tgcctttcag gaggtggcgc
     7081 tgacccacgg tattacgccc cgcatggcgc tcgatcacct cgacggcttt gcgatggacg
     7141 tggctcagac ccgctatgtc acctttgagg atacgctgcg ctactgctat cacgtggcgg
     7201 gcgtggtggg tctgatgatg gccagggtga tgggcgtgcg ggatgagcgg gtgctggatc
     7261 gcgcctgcga tctggggctg gccttccagc tgacgaatat cgcccgggat attattgacg
     7321 atgcggctat tgaccgctgc tatctgcccg ccgagtggct gcaggatgcc gggctgaccc
     7381 cggagaacta tgccgcgcgg gagaatcggg ccgcgctggc gcgggtggcg gagcggctta
     7441 ttgatgccgc agagccgtac tacatctcct cccaggccgg gctacacgat ctgccgccgc
     7501 gctgcgcctg ggcgatcgcc accgcccgca gcgtctaccg ggagatcggt attaaggtaa
     7561 aagcggcggg aggcagcgcc tgggatcgcc gccagcacac cagcaaaggt gaaaaaattg
     7621 ccatgctgat ggcggcaccg gggcaggtta ttcgggcgaa gacgacgagg gtgacgccgc
     7681 gtccggccgg tctttggcag cgtcccgttt aggcgggcgg ccatgacgtt cacgcaggat
     7741 cgcctgtagg tcggcaggct tgcgggcgta aataaaaccg aaggagacgc agccctcccg
     7801 gccgcgcacg ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
     7861 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
     7921 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     7981 catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     8041 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     8101 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     8161 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     8221 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     8281 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     8341 tcattgccat acg
//
