LOCUS       sequence-288147-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of gene coding lycopene bifunctional beta-cyclase for 
            production beta-carotene from lycopene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  TITLE     Engineering E.coli to produce lycopene and its derivations from 
            isopentenol
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6224)
  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: 
            "Unpublished"
COMMENT     Sequence Label: TP2_crtYB
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(326..1408)
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1370..1389
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1409..1486)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1532..1551
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1532..1550
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1551..1575
                     /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)."
     RBS             1590..1612
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3704..3722)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3718..3765
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             4046..4837
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(5164..5181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5298..5843)
                     /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(5378..5397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
       61 caggcttacc cgtcttactg tcgggaattc gcgttggccg attcattaat gcagctggca
      121 cgacaggttt cctctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
      181 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
      241 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
      301 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
      361 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
      421 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
      481 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
      541 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
      601 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
      661 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
      721 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
      781 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
      841 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
      901 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
      961 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     1021 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     1081 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     1141 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     1201 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     1261 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     1321 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     1381 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     1441 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     1501 tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactatagg ggaattgtga
     1561 gcggataaca attcccctgt agaaataatt ttgtttaact ttaagaagga gatatacata
     1621 tgactgcttt ggcctactac caaatccatt tgatctacac cttgccaatt ttgggtttgt
     1681 tgggtttatt gacctctcca attttgacca agttcgacat ctacaagatc tccatcttgg
     1741 ttttcattgc tttctctgct actactccat gggattcttg gattattaga aatggtgctt
     1801 ggacttaccc atctgctgaa tctggtcaag gtgtttttgg tacttttttg gatgtcccat
     1861 acgaagagta cgctttcttc gttattcaaa ccgttattac cggtttggtt tacgttttgg
     1921 ctaccagaca tttgttgcca tctttggctt tgccaaagac tagatcttct gctttgtcat
     1981 tggctttgaa ggctttgatt ccattgccaa tcatctactt gtttaccgct catccatctc
     2041 catcaccaga tccattggtt actgatcatt acttctacat gagagccctg tctttgttga
     2101 ttactccacc aactatgttg ttggctgctt tatctggtga atacgctttt gattggaaat
     2161 ccggtagagc taagtctacc attgctgcta ttatgattcc aaccgtttac ttgatctggg
     2221 ttgattatgt tgctgttggt caagattcct ggtctattaa cgacgaaaaa atcgtcggtt
     2281 ggagattagg tggtgttttg ccaattgaag aagctatgtt cttcctgttg accaacttga
     2341 tgatcgtctt aggtttgtct gcttgtgatc atactcaagc cttgtatttg ttgcatggta
     2401 gaactatcta cggcaacaaa aagatgccat cttctttccc attgatcaca ccaccagttt
     2461 tgtccttgtt tttctcatct agaccatact cttctcagcc aaagagagat ttggaattgg
     2521 ctgttaagtt gctggaagaa aagtccagat cttttttcgt tgcttctgct ggttttccat
     2581 ccgaagttag agaaagattg gttggcttgt atgctttctg tagagttacc gatgatttga
     2641 tcgattcccc agaagtttct tctaatccac atgctaccat cgatatggtt tctgatttct
     2701 tgactttgtt gttcggtcca ccattgcatc catcacaacc agataagatt ttgtcctctc
     2761 cattattgcc accatcacat ccttctagac caactggtat gtatccattg cctccaccac
     2821 catctttgtc tccagctgaa ttggttcaat tcttgactga aagagttcca gtccaatacc
     2881 attttgcttt cagattattg gccaagttgc aaggtttgat cccaagatat ccattggatg
     2941 aattgctgag aggttacacc actgatttga tattcccatt gtctaccgaa gctgttcaag
     3001 ctagaaaaac tccaattgaa actaccgccg atttgttgga ttatggtttg tgtgttgctg
     3061 gttccgttgc tgaattattg gtttatgttt cttgggcttc tgccccatct caagttccag
     3121 ctactattga agaaagagaa gctgttttgg ttgcctctag agaaatgggt actgctttac
     3181 agttggttaa cattgccaga gatattaagg gtgatgctac tgaaggtaga ttctacttgc
     3241 cattgtcatt cttcggtttg agggatgaat ctaagttggc tattccaact gattggactg
     3301 aacctagacc acaagatttc gataagctgt tgtctttgtc accatcttca actttgccat
     3361 cctctaatgc ctctgaatcc tttagatttg agtggaaaac ttactccttg cctttggttg
     3421 cttatgctga agatttggct aaacactcct acaagggtat tgatagattg ccaactgaag
     3481 ttcaagctgg tatgagagct gcttgtgctt cttatttgtt aatcggtaga gaaatcaagg
     3541 tcgtttggaa aggtgatgtt ggtgaaagaa gaactgttgc aggttggaga agagttagaa
     3601 aggttttgtc tgttgttatg tctggttggg aaggtcaatg atgggctgaa gggtttaagg
     3661 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3721 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     3781 atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
     3841 catccagggt gacggtgcct tccctcgact cacacttggg tcgacctgca gaagcttaga
     3901 tctattaccc tgttatccct actcgagttc atgtgcagct ccataagcaa aaggggatga
     3961 taagtttatc accaccgact atttgcaaca gtgccgttga tcgtgctatg atcgactgat
     4021 gtcatcagcg gtggagtgca atgtcatgag ggaagcggtg atcgccgaag tatcgactca
     4081 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     4141 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     4201 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     4261 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     4321 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     4381 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctagc
     4441 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     4501 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     4561 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     4621 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     4681 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     4741 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     4801 cgtgaaaggc gagatcacca aggtagtcgg caaataagat gccgctcgcc agtcgattgg
     4861 ctgagcttat aagttcctat tccgaagttc cgcgaacgcg taaaggatct aggtgaagat
     4921 cctttttgat aatctcatga ccaaaatcct ttaacgttca cacaacatag ccacgggtgt
     4981 tcagctactg acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg
     5041 agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg
     5101 caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc
     5161 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     5221 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     5281 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     5341 cgctcaaatc agtggtgacg aaacccgaca ggactataaa gataccaggc gtttccccct
     5401 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     5461 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     5521 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     5641 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     5701 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     5761 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     5821 cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga
     5881 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     5941 tgtaattctc atgtttgaca gcttatcacc cagtcctgct cgcttcgcta cttggccata
     6001 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     6061 atgtcttcaa acagaaaggc catggggaaa ctacccataa tacaagaaaa gcccgtcacg
     6121 ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt
     6181 cagcagttcc tgccctctga ttttccagtc tgaccacttc ggat
//
