LOCUS       sequence-334291-        8117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PHB production in E. coli (T7-pAC). Please note that this is 
            listed as #140955 in the publication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8117)
  AUTHORS   Liu Y, Low ZJ, Ma X, Liang H, Sinskey AJ, Stephanopoulos G, Zhou K
  TITLE     Using biopolymer bodies for encapsulation of hydrophobic products in
            bacterium.
  JOURNAL   Metab Eng. 2020 Sep;61:206-214. doi: 10.1016/j.ymben.2020.04.006. 
            Epub 2020 Apr 24.
  PUBMED    32339760
REFERENCE   2  (bases 1 to 8117)
  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: "Metab 
            Eng."; date: "2020-09"; volume: "61:206-214. doi"; pages: " 
            10.1016/j.ymben.2020.04.006. Epub 2020 Apr 24"
COMMENT     Sequence Label: pHB-3
FEATURES             Location/Qualifiers
     source          1..8117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(4334..4993)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4908..4927
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4994..5096)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5507..5524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5641..6186)
                     /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(5721..5740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6893..7975)
                     /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     7937..7956
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7976..8053)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(8099..8117,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8099..8117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tggcgaccgg caaaggcgcg gcagcttcca cgcaggaagg caagtcccaa
      121 ccattcaagg tcacgccggg gccattcgat ccagccacat ggctggaatg gtcccgccag
      181 tggcagggca ctgaaggcaa cggccacgcg gccgcgtccg gcattccggg cctggatgcg
      241 ctggcaggcg tcaagatcgc gccggcgcag ctgggtgata tccagcagcg ctacatgaag
      301 gacttctcag cgctgtggca ggccatggcc gagggcaagg ccgaggccac cggtccgctg
      361 cacgaccggc gcttcgccgg cgacgcatgg cgcaccaacc tcccatatcg cttcgctgcc
      421 gcgttctacc tgctcaatgc gcgcgccttg accgagctgg ccgatgccgt cgaggccgat
      481 gccaagaccc gccagcgcat ccgcttcgcg atctcgcaat gggtcgatgc gatgtcgccc
      541 gccaacttcc ttgccaccaa tcccgaggcg cagcgcctgc tgatcgagtc gggcggcgaa
      601 tcgctgcgtg ccggcgtgcg caacatgatg gaagacctga cacgcggcaa gatctcgcag
      661 accgacgaga gcgcgtttga ggtcggccgc aatgtcgcgg tgaccgaagg cgccgtggtc
      721 ttcgagaacg agtacttcca gctgttgcag tacaagccgc tgaccgacaa ggtgcacgcg
      781 cgcccgctgc tgatggtgcc gccgtgcatc aacaagtact acatcctgga cctgcagccg
      841 gagagctcgc tggtgcgcca tgtggtggag cagggacata cggtgtttct ggtgtcgtgg
      901 cgcaatccgg acgccagcat ggccggcagc acctgggacg actacatcga gcacgcggcc
      961 atccgcgcca tcgaagtcgc gcgcgacatc agcggccagg acaagatcaa cgtgctcggc
     1021 ttctgcgtgg gcggcaccat tgtctcgacc gcgctggcgg tgctggccgc gcgcggcgag
     1081 cacccggccg ccagcgtcac gctgctgacc acgctgctgg actttgccga cacgggcatc
     1141 ctcgacgtct ttgtcgacga gggccatgtg cagttgcgcg aggccacgct gggcggcggc
     1201 gccggcgcgc cgtgcgcgct gctgcgcggc cttgagctgg ccaatacctt ctcgttcttg
     1261 cgcccgaacg acctggtgtg gaactacgtg gtcgacaact acctgaaggg caacacgccg
     1321 gtgccgttcg acctgctgtt ctggaacggc gacgccacca acctgccggg gccgtggtac
     1381 tgctggtacc tgcgccacac ctacctgcag aacgagctca aggtaccggg caagctgacc
     1441 gtgtgcggcg tgccggtgga cctggccagc atcgacgtgc cgacctatat ctacggctcg
     1501 cgcgaagacc atatcgtgcc gtggaccgcg gcctatgcct cgaccgcgct gctggcgaac
     1561 aagctgcgct tcgtgctggg tgcgtcgggc catatcgccg gtgtgatcaa cccgccggcc
     1621 aagaacaagc gcagccactg gactaacgat gcgctgccgg agtcgccgca gcaatggctg
     1681 gccggcgcca tcgagcatca cggcagctgg tggccggact ggaccgcatg gctggccggg
     1741 caggccggcg cgaaacgcgc cgcgcccgcc aactatggca atgcgcgcta tcgcgcaatc
     1801 gaacccgcgc ctgggcgata cgtcaaagcc aaggcatgac gcttgcatga gtgccggcgt
     1861 gcgtcatgca cggcgccggc aggcctgcag gttccctccc gtttccattg aaaggactac
     1921 acaatgactg acgttgtcat cgtatccgcc gcccgcaccg cggtcggcaa gtttggcggc
     1981 tcgctggcca agatcccggc accggaactg ggtgccgtgg tcatcaaggc cgcgctggag
     2041 cgcgccggcg tcaagccgga gcaggtgagc gaagtcatca tgggccaggt gctgaccgcc
     2101 ggttcgggcc agaaccccgc acgccaggcc gcgatcaagg ccggcctgcc ggcgatggtg
     2161 ccggccatga ccatcaacaa ggtgtgcggc tcgggcctga aggccgtgat gctggccgcc
     2221 aacgcgatca tggcgggcga cgccgagatc gtggtggccg gcggccagga aaacatgagc
     2281 gccgccccgc acgtgctgcc gggctcgcgc gatggtttcc gcatgggcga tgccaagctg
     2341 gtcgacacca tgatcgtcga cggcctgtgg gacgtgtaca accagtacca catgggcatc
     2401 accgccgaga acgtggccaa ggaatacggc atcacacgcg aggcgcagga tgagttcgcc
     2461 gtcggctcgc agaacaaggc cgaagccgcg cagaaggccg gcaagtttga cgaagagatc
     2521 gtcccggtgc tgatcccgca gcgcaagggc gacccggtgg ccttcaagac cgacgagttc
     2581 gtgcgccagg gcgccacgct ggacagcatg tccggcctca agcccgcctt cgacaaggcc
     2641 ggcacggtga ccgcggccaa cgcctcgggc ctgaacgacg gcgccgccgc ggtggtggtg
     2701 atgtcggcgg ccaaggccaa ggaactgggc ctgaccccgc tggccacgat caagagctat
     2761 gccaacgccg gtgtcgatcc caaggtgatg ggcatgggcc cggtgccggc ctccaagcgc
     2821 gccctgtcgc gcgccgagtg gaccccgcaa gacctggacc tgatggagat caacgaggcc
     2881 tttgccgcgc aggcgctggc ggtgcaccag cagatgggct gggacacctc caaggtcaat
     2941 gtgaacggcg gcgccatcgc catcggccac ccgatcggcg cgtcgggctg ccgtatcctg
     3001 gtgacgctgc tgcacgagat gaagcgccgt gacgcgaaga agggcctggc ctcgctgtgc
     3061 atcggcggcg gcatgggcgt ggcgctggca gtcgagcgca aataaggaag gggttttccg
     3121 gggccgcgcg cggttggcgc ggacccggcg acgataacga agccaatcaa ggagtggaca
     3181 tgactcagcg cattgcgtat gtgaccggcg gcatgggtgg tatcggaacc gccatttgcc
     3241 agcggctggc caaggatggc tttcgtgtgg tggccggttg cggccccaac tcgccgcgcc
     3301 gcgaaaagtg gctggagcag cagaaggccc tgggcttcga tttcattgcc tcggaaggca
     3361 atgtggctga ctgggactcg accaagaccg cattcgacaa ggtcaagtcc gaggtcggcg
     3421 aggttgatgt gctgatcaac aacgccggta tcacccgcga cgtggtgttc cgcaagatga
     3481 cccgcgccga ctgggatgcg gtgatcgaca ccaacctgac ctcgctgttc aacgtcacca
     3541 agcaggtgat cgacggcatg gccgaccgtg gctggggccg catcgtcaac atctcgtcgg
     3601 tgaacgggca gaagggccag ttcggccaga ccaactactc caccgccaag gccggcctgc
     3661 atggcttcac catggcactg gcgcaggaag tggcgaccaa gggcgtgacc gtcaacacgg
     3721 tctctccggg ctatatcgcc accgacatgg tcaaggcgat ccgccaggac gtgctcgaca
     3781 agatcgtcgc gacgatcccg gtcaagcgcc tgggcctgcc ggaagagatc gcctcgatct
     3841 gcgcctggtt gtcgtcggag gagtccggtt tctcgaccgg cgccgacttc tcgctcaacg
     3901 gcggcctgca tatgggctga tgggctgaag ggtttaagcg aagacgaaca ataaggcctc
     3961 ccaaatcggg gggccttttt tattgataac aaaaaggcaa cactatgaca tcggaaaacc
     4021 cgttactggc gctgcgagag aaaatcagcg cgctggatga aaaattatta gcgttactgg
     4081 cagagaggcg cgaattcaaa cagaaaggcc atgggctggc ctcaggcatt tgagaagcac
     4141 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     4201 atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat
     4261 tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac
     4321 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     4381 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     4441 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     4501 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     4561 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     4621 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     4681 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     4741 atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc
     4801 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     4861 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     4921 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     4981 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     5041 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     5101 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     5161 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     5221 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     5281 gtttctatca gctgtccctt tcacacaaca tagccacggg tgttcagcta ctgacggggt
     5341 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     5401 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     5461 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     5521 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     5581 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     5641 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     5701 acgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     5761 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     5821 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     5881 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5941 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     6001 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     6061 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     6121 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     6181 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     6241 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     6301 acagcttatc acccagtcct gctcgcttcg ctacttggcc atacccacgc cgaaacaagc
     6361 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtctt ccctcgactc
     6421 acacttgggg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     6481 tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc
     6541 tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg
     6601 cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcg ggaattcgcg
     6661 ttggccgatt cattaatgca gctggcacga caggtttcct ctagatttca gtgcaattta
     6721 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     6781 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     6841 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     6901 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6961 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     7021 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     7081 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     7141 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     7201 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     7261 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     7321 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     7381 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     7441 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     7501 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     7561 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     7621 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     7681 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     7741 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     7801 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     7861 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     7921 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     7981 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     8041 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta
     8101 atacgactca ctatagg
//
