LOCUS       sequence-109672-        5036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a truncated Arabidopsis beta-carotene hydroxylase cDNA, the
            product of which converts beta-carotene primarily to monocyclic 
            beta-cryptoxanthin in E. coli. Use with pAC-BETA or pAC-BETAipi. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5036)
  AUTHORS   Cunningham FX Jr, Gantt E
  TITLE     A study in scarlet: enzymes of ketocarotenoid biosynthesis in the 
            flowers of Adonis aestivalis.
  JOURNAL   Plant J. 2005 Feb;41(3):478-92.
  PUBMED    15659105
REFERENCE   2  (bases 1 to 5036)
  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: "Plant J."; 
            date: "2005-02"; volume: "41(3)"; pages: "478-92"
COMMENT     Sequence Label: pAtCHYb1-Delta-129N
FEATURES             Location/Qualifiers
     source          1..5036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             446..478
                     /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             join(482..490,491..505)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1219..1236)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1389..1475
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1567..1594
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1614..1705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1706..1774,1775..2566)
                     /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(1924..1943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2737..3325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3226..3245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3479..3496
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3511..3651
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3577..3596)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3737..3759
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3837..3914
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3915..4997
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3934..3953)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcg atggggatct caatgttgga aatgtttggt
      541 acatttgctc tctctgttgg tgctgctgtt ggtatggaat tctgggcaag atgggctcat
      601 agagctctgt ggcacgcttc tctatggaat atgcatgagt cacatcacaa accaagagaa
      661 ggaccgtttg agctaaacga tgtttttgct atagtgaacg ctggtccagc gattggtctc
      721 ctctcttatg gattcttcaa taaaggactc gttcctggtc tctgctttgg cgccgggtta
      781 ggcataacgg tgtttggaat cgcctacatg tttgtccacg atggtctcgt gcacaagcgt
      841 ttccctgtag gtcccatcgc cgacgtccct tacctccgaa aggtcgccgc cgctcaccag
      901 ctacatcaca cagacaagtt caatggtgta ccatatggac tgtttcttgg acccaaggaa
      961 ttggaagaag ttggaggaaa tgaagagtta gataaggaga ttagtcggag aatcaaatca
     1021 tacaaaaagg cctcgggctc cgggtcgagt tcgagttctt gactttaaac aagttttaaa
     1081 tcccaaattc tttttttgtc ttctgtcatt atgatcatct taagacggtc tgcggccgat
     1141 ccgagctcga gatctgcagc tggtaccata tgggaattcg aagcttggct gttttggcgg
     1201 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     1261 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     1321 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     1381 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1441 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     1501 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     1561 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact ctttttgttt
     1621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1681 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1801 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1861 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1921 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1981 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2101 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2281 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2401 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2581 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2641 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2701 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2761 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2821 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2881 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2941 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3001 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3061 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3121 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3181 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3241 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3301 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3361 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3421 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3481 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3541 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3601 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3661 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3721 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3781 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttgaca
     3841 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     3901 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     3961 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     4021 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     4081 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     4141 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     4201 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     4261 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     4321 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     4381 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     4441 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     4501 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     4561 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     4621 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     4681 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     4741 ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat tttcgcctgc
     4801 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     4861 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     4921 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4981 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt gatctg
//
