LOCUS       sequence-334168-        6735 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For PHB production in E. coli (T7-pAC mutant). Please note that this
            is listed as #140954 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 Apr 14, 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-2
FEATURES             Location/Qualifiers
     source          1..6735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(208..225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(342..887)
                     /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(422..441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             1476..1498
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(5770..6429)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6344..6363
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6430..6532)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ttcacacaac atagccacgg gtgttcagct actgacgggg tggtgcgtaa cggcaaaagc
       61 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
      121 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
      181 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
      241 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
      301 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
      361 cctgacaagc atcacgaaat ctgacgctca aatcagtggt gacgaaaccc gacaggacta
      421 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
      481 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
      541 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
      601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
      661 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
      721 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
      781 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
      841 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
      901 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
      961 aagtcagccc catacgatat aagttgtaat tctcatgttt gacagcttat cacccagtcc
     1021 tgctcgcttc gctacttggc catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     1081 cgagcccgat cttccccatc ggtgatgtct tccctcgact cacacttggg agcttttcat
     1141 tctgactgca acgggcaata tgtctctgtg tggattaaaa aaagagtgtc tgatagcagc
     1201 ttctgaactg gttacctgcc gtgagtaaat taaaatttta ttgacttagg tcactaaata
     1261 ctttaaccaa tataggcata gcgcacagac agataaaaat tacagagtac acaacatcca
     1321 tgaaacgcat tagcaccacc attaccacca ccatcaccat taccacaggt aaaggtgcgg
     1381 gctgtcgcgt acaggaaaca cagaaaaaag cccgcacctg acagtgcggg cttttttttc
     1441 gaccaaaggt aacgaggtaa caacctagaa ataattttgt ttaactttaa gaaggagata
     1501 tacatatggc gaccggcaaa ggcgcggcag cttccacgca ggaaggcaag tcccaaccat
     1561 tcaaggtcac gccggggcca ttcgatccag ccacatggct ggaatggtcc cgccagtggc
     1621 agggcactga aggcaacggc cacgcggccg cgtccggcat tccgggcctg gatgcgctgg
     1681 caggcgtcaa gatcgcgccg gcgcagctgg gtgatatcca gcagcgctac atgaaggact
     1741 tctcagcgct gtggcaggcc atggccgagg gcaaggccga ggccaccggt ccgctgcacg
     1801 accggcgctt cgccggcgac gcatggcgca ccaacctccc atatcgcttc gctgccgcgt
     1861 tctacctgct caatgcgcgc gccttgaccg agctggccga tgccgtcgag gccgatgcca
     1921 agacccgcca gcgcatccgc ttcgcgatct cgcaatgggt cgatgcgatg tcgcccgcca
     1981 acttccttgc caccaatccc gaggcgcagc gcctgctgat cgagtcgggc ggcgaatcgc
     2041 tgcgtgccgg cgtgcgcaac atgatggaag acctgacacg cggcaagatc tcgcagaccg
     2101 acgagagcgc gtttgaggtc ggccgcaatg tcgcggtgac cgaaggcgcc gtggtcttcg
     2161 agaacgagta cttccagctg ttgcagtaca agccgctgac cgacaaggtg cacgcgagcc
     2221 cgctgctgat ggtgccgccg tgcatcaaca agtactacat cctggacctg cagccggaga
     2281 gctcgctggt gcgccatgtg gtggagcagg gacatacggt gtttctggtg tcgtggcgca
     2341 atccggacgc cagcatggcc ggcagcacct gggacgacta catcgagcac gcggccatcc
     2401 gcgccatcga agtcgcgcgc gacatcagcg gccaggacaa gatcaacgtg ctcggcttct
     2461 gcgtgggcgg caccattgtc tcgaccgcgc tggcggtgct ggccgcgcgc ggcgagcacc
     2521 cggccgccag cgtcacgctg ctgaccacgc tgctggactt tgccgacacg ggcatcctcg
     2581 acgtctttgt cgacgagggc catgtgcagt tgcgcgaggc cacgctgggc ggcggcgccg
     2641 gcgcgccgtg cgcgctgctg cgcggccttg agctggccaa taccttctcg ttcttgcgcc
     2701 cgaacgacct ggtgtggaac tacgtggtcg acaactacct gaagggcaac acgccggtgc
     2761 cgttcgacct gctgttctgg aacggcgacg ccaccaacct gccggggccg tggtactgct
     2821 ggtacctgcg ccacacctac ctgcagaacg agctcaaggt accgggcaag ctgaccgtgt
     2881 gcggcgtgcc ggtggacctg gccagcatcg acgtgccgac ctatatctac ggctcgcgcg
     2941 aagaccatat cgtgccgtgg accgcggcct atgcctcgac cgcgctgctg gcgaacaagc
     3001 tgcgcttcgt gctgggtgcg tcgggccata tcgccggtgt gatcaacccg ccggccaaga
     3061 acaagcgcag ccactggact aacgatgcgc tgccggagtc gccgcagcaa tggctggccg
     3121 gcgccatcga gcatcacggc agctggtggc cggactggac cgcatggctg gccgggcagg
     3181 ccggcgcgaa acgcgccgcg cccgccaact atggcaatgc gcgctatcgc gcaatcgaac
     3241 ccgcgcctgg gcgatacgtc aaagccaagg catgacgctt gcatgagtgc cggcgtgcgt
     3301 catgcacggc gccggcaggc ctgcaggttc cctcccgttt ccattgaaag gactacacaa
     3361 tgactgacgt tgtcatcgta tccgccgccc gcaccgcggt cggcaagttt ggcggctcgc
     3421 tggccaagat cccggcaccg gaactgggtg ccgtggtcat caaggccgcg ctggagcgcg
     3481 ccggcgtcaa gccggagcag gtgagcgaag tcatcatggg ccaggtgctg accgccggtt
     3541 cgggccagaa ccccgcacgc caggccgcga tcaaggccgg cctgccggcg atggtgccgg
     3601 ccatgaccat caacaaggtg tgcggctcgg gcctgaaggc cgtgatgctg gccgccaacg
     3661 cgatcatggc gggcgacgcc gagatcgtgg tggccggcgg ccaggaaaac atgagcgccg
     3721 ccccgcacgt gctgccgggc tcgcgcgatg gtttccgcat gggcgatgcc aagctggtcg
     3781 acaccatgat cgtcgacggc ctgtgggacg tgtacaacca gtaccacatg ggcatcaccg
     3841 ccgagaacgt ggccaaggaa tacggcatca cacgcgaggc gcaggatgag ttcgccgtcg
     3901 gctcgcagaa caaggccgaa gccgcgcaga aggccggcaa gtttgacgaa gagatcgtcc
     3961 cggtgctgat cccgcagcgc aagggcgacc cggtggcctt caagaccgac gagttcgtgc
     4021 gccagggcgc cacgctggac agcatgtccg gcctcaagcc cgccttcgac aaggccggca
     4081 cggtgaccgc ggccaacgcc tcgggcctga acgacggcgc cgccgcggtg gtggtgatgt
     4141 cggcggccaa ggccaaggaa ctgggcctga ccccgctggc cacgatcaag agctatgcca
     4201 acgccggtgt cgatcccaag gtgatgggca tgggcccggt gccggcctcc aagcgcgccc
     4261 tgtcgcgcgc cgagtggacc ccgcaagacc tggacctgat ggagatcaac gaggcctttg
     4321 ccgcgcaggc gctggcggtg caccagcaga tgggctggga cacctccaag gtcaatgtga
     4381 acggcggcgc catcgccatc ggccacccga tcggcgcgtc gggctgccgt atcctggtga
     4441 cgctgctgca cgagatgaag cgccgtgacg cgaagaaggg cctggcctcg ctgtgcatcg
     4501 gcggcggcat gggcgtggcg ctggcagtcg agcgcaaata aggaaggggt tttccggggc
     4561 cgcgcgcggt tggcgcggac ccggcgacga taacgaagcc aatcaaggag tggacatgac
     4621 tcagcgcatt gcgtatgtga ccggcggcat gggtggtatc ggaaccgcca tttgccagcg
     4681 gctggccaag gatggctttc gtgtggtggc cggttgcggc cccaactcgc cgcgccgcga
     4741 aaagtggctg gagcagcaga aggccctggg cttcgatttc attgcctcgg aaggcaatgt
     4801 ggctgactgg gactcgacca agaccgcatt cgacaaggtc aagtccgagg tcggcgaggt
     4861 tgatgtgctg atcaacaacg ccggtatcac ccgcgacgtg gtgttccgca agatgacccg
     4921 cgccgactgg gatgcggtga tcgacaccaa cctgacctcg ctgttcaacg tcaccaagca
     4981 ggtgatcgac ggcatggccg accgtggctg gggccgcatc gtcaacatct cgtcggtgaa
     5041 cgggcagaag ggccagttcg gccagaccaa ctactccacc gccaaggccg gcctgcatgg
     5101 cttcaccatg gcactggcgc aggaagtggc gaccaagggc gtgaccgtca acacggtctc
     5161 tccgggctat atcgccaccg acatggtcaa ggcgatccgc caggacgtgc tcgacaagat
     5221 cgtcgcgacg atcccggtca agcgcctggg cctgccggaa gagatcgcct cgatctgcgc
     5281 ctggttgtcg tcggaggagt ccggtttctc gaccggcgcc gacttctcgc tcaacggcgg
     5341 cctgcatatg ggctgatggg ctgaagggtt taagcgaaga cgaacaataa ggcctcccaa
     5401 atcggggggc cttttttatt gataacaaaa aggcaacact atgacatcgg aaaacccgtt
     5461 actggcgctg cgagagaaaa tcagcgcgct ggatgaaaaa ttattagcgt tactggcaga
     5521 gaggcgcgaa ttcaaacaga aaggccatgg gctggcctca ggcatttgag aagcacacgg
     5581 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     5641 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
     5701 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
     5761 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     5821 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     5881 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     5941 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     6001 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     6061 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     6121 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     6181 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     6241 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     6301 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     6361 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     6421 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     6481 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     6541 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     6601 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
     6661 tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt
     6721 ctatcagctg tccct
//
