LOCUS       sequence-83479-e        4326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces an active HBFD (carotenoid 4-hydroxy-beta-ring 
            4-dehydrogenase) enzyme in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4326)
  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 4326)
  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: pHBFD1
FEATURES             Location/Qualifiers
     source          1..4326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2223..2239
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2223..2239
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3342..3353
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3682..3701)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3683..3701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3708..3725)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3708..3724)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3717..3739)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3865..4320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4002..4023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4214..4233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctcgaaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     2221 gctctagaac tagtggatcc cccgggctgc aggaattcgg cacgaggaag aacattacat
     2281 ggctcctgtt ctccttggat tgaaaccaac tctctccact ggaagcgtcg tcaaagagac
     2341 taatgtagga agcacacttg ctagtcccct taacaaaacc cagaattcaa gggttttggt
     2401 tttgggcgga acagggaagg tcggtggttc cacagctttg gctctctcca agttctcacc
     2461 tgacctcagg cttgtgattg gaggtcgaaa cagggagaaa ggtgatgctg tagtgtctaa
     2521 actaggagaa aactccgagt ttgttgaagt caacgttgac agtgtgagat ctttagaatc
     2581 tgctctcgaa gatgtggacc ttgtagttca tgcagctgga ccttttcaac aagcggagaa
     2641 gtgcactgtt ctagaagctg caatatctac caggacggcc tatgtggatg tatgtgataa
     2701 tacaagttat tccatgcaag caaagtcttt tcatgataaa gcagtggctg ccaacgttcc
     2761 tgccataaca actgctggaa ttttccctgg agtgagcaat gtgatagcag ctgagctagt
     2821 gcgatcagca agagatgaaa acactgaacc tcaaagacta agattctcct attttaccgc
     2881 gggttctggt ggtgctggtc caacgtcgtt agttactagc ttcttgcttc ttggtgaaga
     2941 ggttgttgct tacagtgaag gcgaaaaagt cgaattaaag ccttatacag ggaagcttaa
     3001 cattgacttc gggaagggag ttgggaaaag agacgtttat ttgtggaact tgccggaagt
     3061 aagaagtggt catgagatct taggagtacc aactgtgagt gctcgattcg gtactgcacc
     3121 tttcttctgg aattgggcga tggtagctat gacaactctc cttcctcctg gtattctgag
     3181 agacagaaat aaaatcggaa tgttggcaaa ttttgtgtac ccttctgtac aaatttttga
     3241 tgggattgca ggagaatgtc ttgcaatgcg ggttgattta gagtgcgcaa atgggcgcaa
     3301 tacttttggt atactcagtc atgaacgtct ctctgtatta gtgggaactt caactgcggt
     3361 gtttgctatg gcaattcttg aaggaagtac gcagcctgga gtttggtttc cagaagagcc
     3421 tggagggatt gcaataagtg acagagagtt acttctacaa cgagcatcac aaggagcgat
     3481 taacttcatt atgaagcagt agagtaatag attggattat tcattatgta gcccagaatg
     3541 acattattta catgtaatgt tgcttctatg tatcaataac ataaatcaca agtcattcgt
     3601 atttatataa gtattcagtc catatctggg agcaaaaaaa aaaaaaaaaa aaactcgagg
     3661 gggggcccgg tacccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
     3721 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3781 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3841 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     3901 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3961 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4021 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4081 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4141 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4201 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4261 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4321 caggtg
//
