LOCUS       sequence-334294-        6735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PHB production in E. coli (thrC3-pAC). Please note that this is 
            listed as #140956 in the publication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6735)
  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 6735)
  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-4
FEATURES             Location/Qualifiers
     source          1..6735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(4295..4954)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4869..4888
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4955..5057)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5468..5485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5602..6147)
                     /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(5682..5701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atggcgaccg gcaaaggcgc ggcagcttcc
       61 acgcaggaag gcaagtccca accattcaag gtcacgccgg ggccattcga tccagccaca
      121 tggctggaat ggtcccgcca gtggcagggc actgaaggca acggccacgc ggccgcgtcc
      181 ggcattccgg gcctggatgc gctggcaggc gtcaagatcg cgccggcgca gctgggtgat
      241 atccagcagc gctacatgaa ggacttctca gcgctgtggc aggccatggc cgagggcaag
      301 gccgaggcca ccggtccgct gcacgaccgg cgcttcgccg gcgacgcatg gcgcaccaac
      361 ctcccatatc gcttcgctgc cgcgttctac ctgctcaatg cgcgcgcctt gaccgagctg
      421 gccgatgccg tcgaggccga tgccaagacc cgccagcgca tccgcttcgc gatctcgcaa
      481 tgggtcgatg cgatgtcgcc cgccaacttc cttgccacca atcccgaggc gcagcgcctg
      541 ctgatcgagt cgggcggcga atcgctgcgt gccggcgtgc gcaacatgat ggaagacctg
      601 acacgcggca agatctcgca gaccgacgag agcgcgtttg aggtcggccg caatgtcgcg
      661 gtgaccgaag gcgccgtggt cttcgagaac gagtacttcc agctgttgca gtacaagccg
      721 ctgaccgaca aggtgcacgc gcgcccgctg ctgatggtgc cgccgtgcat caacaagtac
      781 tacatcctgg acctgcagcc ggagagctcg ctggtgcgcc atgtggtgga gcagggacat
      841 acggtgtttc tggtgtcgtg gcgcaatccg gacgccagca tggccggcag cacctgggac
      901 gactacatcg agcacgcggc catccgcgcc atcgaagtcg cgcgcgacat cagcggccag
      961 gacaagatca acgtgctcgg cttctgcgtg ggcggcacca ttgtctcgac cgcgctggcg
     1021 gtgctggccg cgcgcggcga gcacccggcc gccagcgtca cgctgctgac cacgctgctg
     1081 gactttgccg acacgggcat cctcgacgtc tttgtcgacg agggccatgt gcagttgcgc
     1141 gaggccacgc tgggcggcgg cgccggcgcg ccgtgcgcgc tgctgcgcgg ccttgagctg
     1201 gccaatacct tctcgttctt gcgcccgaac gacctggtgt ggaactacgt ggtcgacaac
     1261 tacctgaagg gcaacacgcc ggtgccgttc gacctgctgt tctggaacgg cgacgccacc
     1321 aacctgccgg ggccgtggta ctgctggtac ctgcgccaca cctacctgca gaacgagctc
     1381 aaggtaccgg gcaagctgac cgtgtgcggc gtgccggtgg acctggccag catcgacgtg
     1441 ccgacctata tctacggctc gcgcgaagac catatcgtgc cgtggaccgc ggcctatgcc
     1501 tcgaccgcgc tgctggcgaa caagctgcgc ttcgtgctgg gtgcgtcggg ccatatcgcc
     1561 ggtgtgatca acccgccggc caagaacaag cgcagccact ggactaacga tgcgctgccg
     1621 gagtcgccgc agcaatggct ggccggcgcc atcgagcatc acggcagctg gtggccggac
     1681 tggaccgcat ggctggccgg gcaggccggc gcgaaacgcg ccgcgcccgc caactatggc
     1741 aatgcgcgct atcgcgcaat cgaacccgcg cctgggcgat acgtcaaagc caaggcatga
     1801 cgcttgcatg agtgccggcg tgcgtcatgc acggcgccgg caggcctgca ggttccctcc
     1861 cgtttccatt gaaaggacta cacaatgact gacgttgtca tcgtatccgc cgcccgcacc
     1921 gcggtcggca agtttggcgg ctcgctggcc aagatcccgg caccggaact gggtgccgtg
     1981 gtcatcaagg ccgcgctgga gcgcgccggc gtcaagccgg agcaggtgag cgaagtcatc
     2041 atgggccagg tgctgaccgc cggttcgggc cagaaccccg cacgccaggc cgcgatcaag
     2101 gccggcctgc cggcgatggt gccggccatg accatcaaca aggtgtgcgg ctcgggcctg
     2161 aaggccgtga tgctggccgc caacgcgatc atggcgggcg acgccgagat cgtggtggcc
     2221 ggcggccagg aaaacatgag cgccgccccg cacgtgctgc cgggctcgcg cgatggtttc
     2281 cgcatgggcg atgccaagct ggtcgacacc atgatcgtcg acggcctgtg ggacgtgtac
     2341 aaccagtacc acatgggcat caccgccgag aacgtggcca aggaatacgg catcacacgc
     2401 gaggcgcagg atgagttcgc cgtcggctcg cagaacaagg ccgaagccgc gcagaaggcc
     2461 ggcaagtttg acgaagagat cgtcccggtg ctgatcccgc agcgcaaggg cgacccggtg
     2521 gccttcaaga ccgacgagtt cgtgcgccag ggcgccacgc tggacagcat gtccggcctc
     2581 aagcccgcct tcgacaaggc cggcacggtg accgcggcca acgcctcggg cctgaacgac
     2641 ggcgccgccg cggtggtggt gatgtcggcg gccaaggcca aggaactggg cctgaccccg
     2701 ctggccacga tcaagagcta tgccaacgcc ggtgtcgatc ccaaggtgat gggcatgggc
     2761 ccggtgccgg cctccaagcg cgccctgtcg cgcgccgagt ggaccccgca agacctggac
     2821 ctgatggaga tcaacgaggc ctttgccgcg caggcgctgg cggtgcacca gcagatgggc
     2881 tgggacacct ccaaggtcaa tgtgaacggc ggcgccatcg ccatcggcca cccgatcggc
     2941 gcgtcgggct gccgtatcct ggtgacgctg ctgcacgaga tgaagcgccg tgacgcgaag
     3001 aagggcctgg cctcgctgtg catcggcggc ggcatgggcg tggcgctggc agtcgagcgc
     3061 aaataaggaa ggggttttcc ggggccgcgc gcggttggcg cggacccggc gacgataacg
     3121 aagccaatca aggagtggac atgactcagc gcattgcgta tgtgaccggc ggcatgggtg
     3181 gtatcggaac cgccatttgc cagcggctgg ccaaggatgg ctttcgtgtg gtggccggtt
     3241 gcggccccaa ctcgccgcgc cgcgaaaagt ggctggagca gcagaaggcc ctgggcttcg
     3301 atttcattgc ctcggaaggc aatgtggctg actgggactc gaccaagacc gcattcgaca
     3361 aggtcaagtc cgaggtcggc gaggttgatg tgctgatcaa caacgccggt atcacccgcg
     3421 acgtggtgtt ccgcaagatg acccgcgccg actgggatgc ggtgatcgac accaacctga
     3481 cctcgctgtt caacgtcacc aagcaggtga tcgacggcat ggccgaccgt ggctggggcc
     3541 gcatcgtcaa catctcgtcg gtgaacgggc agaagggcca gttcggccag accaactact
     3601 ccaccgccaa ggccggcctg catggcttca ccatggcact ggcgcaggaa gtggcgacca
     3661 agggcgtgac cgtcaacacg gtctctccgg gctatatcgc caccgacatg gtcaaggcga
     3721 tccgccagga cgtgctcgac aagatcgtcg cgacgatccc ggtcaagcgc ctgggcctgc
     3781 cggaagagat cgcctcgatc tgcgcctggt tgtcgtcgga ggagtccggt ttctcgaccg
     3841 gcgccgactt ctcgctcaac ggcggcctgc atatgggctg atgggctgaa gggtttaagc
     3901 gaagacgaac aataaggcct cccaaatcgg ggggcctttt ttattgataa caaaaaggca
     3961 acactatgac atcggaaaac ccgttactgg cgctgcgaga gaaaatcagc gcgctggatg
     4021 aaaaattatt agcgttactg gcagagaggc gcgaattcaa acagaaaggc catgggctgg
     4081 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     4141 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     4201 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag
     4261 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     4321 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     4381 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     4441 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     4501 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     4561 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     4621 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     4681 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt
     4741 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     4801 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     4861 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     4921 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     4981 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     5041 ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat
     5101 caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt
     5161 cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt
     5221 tgaggtgctc cagtggcttc tgtttctatc agctgtccct ttcacacaac atagccacgg
     5281 gtgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta
     5341 gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat
     5401 gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt
     5461 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     5521 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     5581 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     5641 ctgacgctca aatcagtggt gacgaaaccc gacaggacta taaagatacc aggcgtttcc
     5701 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     5761 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     5821 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     5881 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     5941 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     6001 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     6061 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     6121 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
     6181 tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat
     6241 aagttgtaat tctcatgttt gacagcttat cacccagtcc tgctcgcttc gctacttggc
     6301 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     6361 ggtgatgtct tccctcgact cacacttggg agcttttcat tctgactgca acgggcaata
     6421 tgtctctgtg tggattaaaa aaagagtgtc tgatagcagc ttctgaactg gttacctgcc
     6481 gtgagtaaat taaaatttta ttgacttagg tcactaaata ctttaaccaa tataggcata
     6541 gcgcacagac agataaaaat tacagagtac acaacatcca tgaaacgcat tagcaccacc
     6601 attaccacca ccatcaccat taccacaggt aacggtgcgg gctgacgcgt acaggaaaca
     6661 cagaaaaaag cccgcacctg acagtgcggg cttttttttc gaccaaaggt aacgaggtaa
     6721 caacctagaa ataat
//
