LOCUS       sequence-83480-e        6707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Will convert beta-carotene into astaxanthin in E. coli when 
            expression of the hbfd1 cDNA is induced with arabinose. Use with 
            pAC-BETA or pAC-BETAipi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6707)
  AUTHORS   Cunningham FX Jr, Gantt E
  TITLE     Elucidation of the pathway to astaxanthin in the flowers of Adonis 
            aestivalis.
  JOURNAL   Plant Cell. 2011 Aug;23(8):3055-69. doi: 10.1105/tpc.111.086827. 
            Epub 2011 Aug 23.
  PUBMED    21862704
REFERENCE   2  (bases 1 to 6707)
  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: 
            "10.1105/tpc.111.086827"; journalName: "Plant Cell"; date: 
            "2011-08"; volume: "23"; issue: "8"; pages: "3055-69"
COMMENT     Sequence Label: pCBFD1/HBFD1Bad
FEATURES             Location/Qualifiers
     source          1..6707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1382..1393
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2798..2814)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2814)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2851..2869)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2890..2906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2890..2906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2903..2925)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2914..2930
                     /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)."
     promoter        complement(join(2938..2944,2945..2962,2963..2968))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2983..3004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3108..3125)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3278..3364
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3456..3483
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3502..3593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3594..3662,3663..4454)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3812..3831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4625..5213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5114..5133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5367..5384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5399..5539
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5465..5484)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5625..5647
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5803..6681)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggctcctgtt ctccttggat tgaaaccaac tctctccact
      361 ggaagcgtcg tcaaagagac taatgtagga agcacacttg ctagtcccct taacaaaacc
      421 cagaattcaa gggttttggt tttgggcgga acagggaagg tcggtggttc cacagctttg
      481 gctctctcca agttctcacc tgacctcagg cttgtgattg gaggtcgaaa cagggagaaa
      541 ggtgatgctg tagtgtctaa actaggagaa aactccgagt ttgttgaagt caacgttgac
      601 agtgtgagat ctttagaatc tgctctcgaa gatgtggacc ttgtagttca tgcagctgga
      661 ccttttcaac aagcggagaa gtgcactgtt ctagaagctg caatatctac caggacggcc
      721 tatgtggatg tatgtgataa tacaagttat tccatgcaag caaagtcttt tcatgataaa
      781 gcagtggctg ccaacgttcc tgccataaca actgctggaa ttttccctgg agtgagcaat
      841 gtgatagcag ctgagctagt gcgatcagca agagatgaaa acactgaacc tcaaagacta
      901 agattctcct attttaccgc gggttctggt ggtgctggtc caacgtcgtt agttactagc
      961 ttcttgcttc ttggtgaaga ggttgttgct tacagtgaag gcgaaaaagt cgaattaaag
     1021 ccttatacag ggaagcttaa cattgacttc gggaagggag ttgggaaaag agacgtttat
     1081 ttgtggaact tgccggaagt aagaagtggt catgagatct taggagtacc aactgtgagt
     1141 gctcgattcg gtactgcacc tttcttctgg aattgggcga tggtagctat gacaactctc
     1201 cttcctcctg gtattctgag agacagaaat aaaatcggaa tgttggcaaa ttttgtgtac
     1261 ccttctgtac aaatttttga tgggattgca ggagaatgtc ttgcaatgcg ggttgattta
     1321 gagtgcgcaa atgggcgcaa tacttttggt atactcagtc atgaacgtct ctctgtatta
     1381 gtgggaactt caactgcggt gtttgctatg gcaattcttg aaggaagtac gcagcctgga
     1441 gtttggtttc cagaagagcc tggagggatt gcaataagtg acagagagtt acttctacaa
     1501 cgagcatcac aaggagcgat taacttcatt atgaagcagt agagtaatag attggattat
     1561 tcattatgta gcccagatct gaattcgtcg acaagcttct cgagtttttt ttgctttaat
     1621 taatatatat tgatattgat caccagataa aactacaaac tctaattttt atttttaata
     1681 atggaattcc acaacaatag atttggcctt tgtaaccgta actccgcgac ctatacatca
     1741 aaaaaaaatt ctaaaaaaga gtatggtaac tttgaattac tacaacgaca caaatgtgtg
     1801 acttgtaaca ccgataaact aaaacctcat aaatgtaaaa gagttgattc atgtagatga
     1861 ttgcgttcgt ttagctgtac gactgatcac cctctccaat tcttcagtgc ctcctcttac
     1921 ttcttccaat tcctggggtc caaggaacag gccaaatggt acaccctgaa attttcctga
     1981 gtgatggatt tggtgagctg cagccagctt gtggaaataa gggacgtttg caataagccc
     2041 tactgggaac cttcggtgtg aaaggccatt gtgaagaaaa atgtaagcca tgccacagac
     2101 tgtcgttcca agaccggtac caaagcaggc tccaggaagg aggccttcat ttgagaatcc
     2161 ataattgata agagcaatag caggaagcgc gttaataata gcaaacacat cattgaactc
     2221 gaagcggcct tttcgtgacc tatggtgaga cttgtgaatg taccacaaag aatcgtgcca
     2281 gagttctttg tgaacccatc ctgacaaaaa ttccgtgcca atcgcagcag caacaaaggc
     2341 ggcaaacgta atccccattt ctgcgacagg aatatctcca cactccatta gccgtgaaaa
     2401 cttgaagtaa acagcaggtc cgatcattga aaggatccca agacaagata cgggtgcgac
     2461 aatggagcca gtggaccgtc ttgattgttt cccccctagc gttttcttta tatgaattat
     2521 gccagaatcc gtctgttcta ttagttcttc ctcgttctct tcctcttctt caatttgagg
     2581 aatatcaagg ttccttgttc tctctgcaac agagcagata catgcagcac gatgtgtctt
     2641 tttctttctc ataggcgact ccaccacaac tggagagaag agcaaacatg ggcggttgag
     2701 aatgtcttgt tttgagtgca acaagagatt cttgtggaaa gaataacttg tactgaacac
     2761 tgagattgct ctcgtgccga attcctgcag cccgggggat ccactagttc tagagcggcc
     2821 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattt cgagcttggc
     2881 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2941 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3001 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcatat
     3061 gggaattcga agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     3121 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     3181 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     3241 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     3301 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     3361 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     3421 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     3481 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
     3541 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3601 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3661 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3721 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3781 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     3841 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3901 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3961 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4021 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4081 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4141 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4201 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4261 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4321 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4381 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4441 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4501 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4561 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4621 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4681 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4741 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4801 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4861 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4921 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4981 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5041 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5101 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5161 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5221 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5281 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5341 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5401 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     5461 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     5521 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5581 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5641 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
     5701 ggcgaagcgg catgcataat gtgcctgtca aatggacgaa gcagggattc tgcaaaccct
     5761 atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca acttgacggc
     5821 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
     5881 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
     5941 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
     6001 gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg
     6061 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
     6121 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
     6181 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
     6241 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
     6301 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
     6361 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
     6421 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
     6481 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
     6541 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
     6601 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
     6661 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagag
//
