LOCUS       sequence-106545-        7240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses crtEBI genes for neurosporene production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7240)
  AUTHORS   Farasat I, Kushwaha M, Collens J, Easterbrook M, Guido M, Salis HM
  TITLE     Efficient search, mapping, and optimization of multi-protein genetic
            systems in diverse bacteria.
  JOURNAL   Mol Syst Biol. 2014 Jun 21;10:731. doi: 10.15252/msb.20134955.
  PUBMED    24952589
REFERENCE   2  (bases 1 to 7240)
  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: "Mol Syst 
            Biol."; date: "2014-06-21"; volume: "10:731. doi"; pages: " 
            10.15252/msb.20134955"
COMMENT     Sequence Label: LacI_PlacO1_crtEBI_13
FEATURES             Location/Qualifiers
     source          1..7240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    323..344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(357..1439)
                     /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     1401..1420
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1440..1517)
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    1581..1597
                     /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)."
     protein_bind    1604..1620
                     /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             1675..1683
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             3687..3695
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      complement(5406..5433)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5449..5520)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             5633..6292
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5699..5718)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      6313..6407
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6496..7084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6985..7004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccctcg
       61 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
      121 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
      181 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
      241 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaagt
      301 tatgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      361 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      421 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
      481 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
      541 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
      601 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
      661 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
      721 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
      781 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
      841 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
      901 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
      961 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     1021 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     1081 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     1141 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     1201 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     1261 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     1321 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     1381 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     1441 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     1501 cgccattcga tggtgtcaac gtaaatgcat gccgcttcgc cttccggcca ccagaatagc
     1561 ctgcgattca acgtcgacaa ttgtgagcgg ataacaattg acattgtgag cggataacaa
     1621 gatactgagc acatcagcag gacgcactga ccgaattcga tttttaggaa cagttaagga
     1681 ggttaataat ggcattcgaa caacgtattg aagctgctat ggctgctgct attgcacgtg
     1741 gccagggtag cgaagctccg tccaaactgg caactgcgct ggactacgct gtcacccctg
     1801 gtggtgcgcg cattcgtccg actctgctgc tgtctgtcgc caccgcttgc ggcgatgacc
     1861 gtcctgccct gagcgacgcg gctgcggttg ccctggaact gatccattgc gcttctctgg
     1921 tgcatgacga cctgccgtgc ttcgacgacg ctgaaatccg tcgtggtaag ccaaccgttc
     1981 atcgtgcata cagcgaaccg ctggctatcc tgactggtga ttccctgatc gttatgggtt
     2041 tcgaagttct ggcgcgtgcg gcggctgatc agccacaacg tgccctgcaa ctggtaacgg
     2101 ctctggcggt acgcactggt atgccgatgg gcatctgcgc tggtcagggt tgggaaagcg
     2161 aatctcagat caacctgtct gcataccacc gtgccaaaac tggtgccctg ttcatcgcgg
     2221 ctactcagat gggcgcaatc gcagcgggtt acgaggcgga accgtgggaa gaactgggtg
     2281 cgcgtatcgg cgaagcgttc caggttgcag atgatctgcg tgacgcgctg tgcgatgccg
     2341 aaaccctggg taagccggca ggtcaggacg aaatccatgc acgtccgaac gctgttcgcg
     2401 agtatggtgt agaaggcgcg gctaaacgcc tgaaagacat cctgggtggc gcgatcgcat
     2461 ccatcccgtc ctgtccaggt gaagcaatgc tggcagagat ggttcgtcgc tacgccgaga
     2521 aaatcgttcc agctcaagtt gctgcacgcg tgtaagagta caatagaaat taaaatcagg
     2581 aggtcaacaa tgattgcttc tgctgatctg gatgcgtgtc gtgaaatgat tcgtaccggc
     2641 tcctatagct ttcatgccgc ttctcgtctg ctgccggaac gcgtacgtgc gccgagcctg
     2701 gcgctgtacg ccttctgccg tgtagctgac gatgctgtgg atgaagctgt taacgatggc
     2761 cagcgtgaag aagatgctga agtgaaacgc cgcgcagtac tgagcctgcg tgatcgtctg
     2821 gatctggtgt atggcggccg tccgcgtaac gctcctgctg accgcgcctt tgcggcggtc
     2881 gtcgaagaat ttgaaatgcc gcgtgctctg ccggaagcgc tgctggaagg tctggcgtgg
     2941 gatgcggtag gccgttccta cgactccttc agcggtgttc tggattactc tgcgcgtgtt
     3001 gccgcagccg ttggcgcgat gatgtgtgtt ctgatgcgcg ttcgcgatcc tgatgtcctg
     3061 gctcgtgcgt gcgatctggg tctggcgatg caactgacca acatcgctcg tgacgttggc
     3121 actgacgccc gttctggtcg catctacctg ccgcgcgact ggatggaaga ggaaggcctg
     3181 ccggtggaag agttcctggc acgcccggtg gttgatgatc gcatccgtgc agtcactcac
     3241 cgcctgctgc gcgctgcgga ccgtctgtac ctgcgttctg aggccggtgt gtgtggcctg
     3301 ccgctggctt gtcgtccggg catttacgcg gcacgtcaca tctatgctgg tattggtgat
     3361 gaaatcgcgc gtaacggtta tgattctgtt acccgtcgcg ctttcaccac tcgtcgccag
     3421 aaactggttt ggctgggtct gtccgctact cgcgctgctc tgagcccatt cggtccaggt
     3481 tgtgcaaccc tgcacgccgc tccggaacca gaagtcgcat tcctggttaa cgcggcagca
     3541 cgcgctcgtc ctcagcgtgg ccgtagcgaa gctctgatta gcgttctggc tcaactggag
     3601 gctcaggatc gtcagatcag ccgccagcgt ctgggtaacc gtgcgaaccc gatctaactc
     3661 gaagcttaga ttttaaataa caatactaag gaggtgcaac atgccatcta tttctccagc
     3721 ctctgacgct gatcgtgctc tggttatcgg ctctggcctg ggcggcctgg ccgctgctat
     3781 gcgtctgggt gcaaaaggtt ggcgtgttac cgtaatcgat aaactggacg ttccgggtgg
     3841 ccgtggctcc agcatcacgc aggagggcca tcgttttgac ctgggcccga ccattgttac
     3901 tgtacctcaa agcctgcgtg acctgtggaa aacttgcggt cgcgacttcg acgcagatgt
     3961 ggagctgaaa ccgattgacc cattctacga agttcgttgg ccagatggct ctcactttac
     4021 cgttcgtcag tctactgaag ctatgaaagc tgaagtagca cgcctgtctc caggcgatgt
     4081 tgcgggctac gagaaattcc tgaaggattc cgagaaacgt tattggttcg gttatgagga
     4141 cctgggtcgt cgctccatgc acaaactgtg ggatctgatc aaagtcctgc cgaccttcgg
     4201 catgatgcgt gctgatcgtt ccgtctacca gcacgcggcc ctgcgcgtca aggatgaacg
     4261 cctgcgcatg gcgctgtcct tccatcctct gttcattggt ggtgatccgt tcaatgtgac
     4321 cagcatgtac atcctggtat ctcaactgga gaaagaattt ggtgtacatt acgcgatcgg
     4381 cggtgtggcc gccatcgcag ccgcgatggc gaaagtgatt gaaggtcagg gcggctcctt
     4441 ccgcatgaac actgaagtgg acgaaatcct ggttgaaaaa ggcaccgcta ccggtgttcg
     4501 cctggcgtct ggcgaagtac tgcgcgctgg tctggttgta agcaatgcgg acgcaggcca
     4561 tacctacatg cgtctgctgc gcaatcaccc tcgccgtcgt tggactgacg cccacgttaa
     4621 atctcgccgc tggagcatgg gcctgtttgt ttggtacttt ggcaccaaag gtactaaagg
     4681 catgtggccg gatgtgggtc atcacaccat cgtgaacgct ccgcgttaca aaggcctggt
     4741 tgaggacatc ttcctgaaag gtaaactggc caaagacatg tccctgtaca tccatcgtcc
     4801 atctatcact gatccgaccg ttgcgccaga aggcgacgat accttctacg cgctgtcccc
     4861 agtgccgcac ctgaaacagg cacagccggt agactggcag gcggttgcgg agccgtaccg
     4921 cgaaagcgtc ctggaagtcc tggaacaatc catgccgggc attggtgaac gcattggtcc
     4981 aagcctggtg ttcacgccag aaactttccg tgatcgttac ctgtccccgt ggggtgcagg
     5041 cttctctatc gaaccgcgca tcctgcaatc tgcttggttc cgtccgcata atatctctga
     5101 agaagtcgca aatctgtttc tggttggtgc gggcacccac ccaggtgccg gtgtaccggg
     5161 tgttattggc tctgcggagg tcatggcgaa actggctccg gatgcgccgc gtgcgcgccg
     5221 tgaagctgaa ccggctgagc gcctggcagc agaataacca taacacacac acacacaagg
     5281 tcgtcacagc accggtgctt aataaagatc ttcgacgtct gctgtgtaaa aaaaaacccc
     5341 gcccctgaca gggcggggtt ttttttcgcg gttgtgaatg ccatatggct gtgctgtggt
     5401 ataaacgcag aaaggcccac ccgaaggtga gccagtgtga ctctagtaga gagcgttcac
     5461 cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg
     5521 atgcctggcc taggttcacg taagaggttc caactttcac cataatgaaa taagatcact
     5581 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     5641 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     5701 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     5761 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     5821 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
     5881 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     5941 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     6001 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     6061 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     6121 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     6181 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     6241 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aatttgatat
     6301 cggtaccgct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     6361 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agcacccggg
     6421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6601 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7201 agctgatacc tggatcctga taaatgattt ctgcagaaaa
//
