LOCUS       sequence-420764-        7906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PHB production in E. coli (PT7-pMB1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  TITLE     Harnessing xylitol utilization in Escherichia coli for 
            antibiotic-free plasmid and recombinant protein production.
REFERENCE   2  (bases 1 to 7906)
  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"
COMMENT     Sequence Label: PHB1
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      358..946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     847..866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1089..1106
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1667..2749)
                     /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     2711..2730
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2750..2827)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2873..2892
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2873..2891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2892..2916
                     /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             2931..2953
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(7225..7884)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7799..7818
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7885..7906,1..81))
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
       61 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg
      121 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt
      181 tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg tatgatggtg
      241 tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctttcaca caacatagcc
      301 acgggagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
      361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
      421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
      481 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
      541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
      601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
      661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
      721 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
      781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      961 ggccttttta cggttcctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1021 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1081 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     1141 ttacgcatct gtgcggtatt tcacaccgca tatgctggat ccttccctcg actcacactt
     1201 ggggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc
     1261 gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt
     1321 tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca
     1381 gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtcgggaatt cgcgttggcc
     1441 gattcattaa tgcagctggc acgacaggtt tcctctagat ttcagtgcaa tttatctctt
     1501 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat
     1561 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     1621 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     1681 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1741 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     1801 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     1861 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     1921 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
     1981 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     2041 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     2101 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     2161 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     2221 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     2281 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     2341 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     2401 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     2461 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     2521 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     2581 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     2641 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     2701 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     2761 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     2821 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga
     2881 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     2941 tttaagaagg agatatacat atggcgaccg gcaaaggcgc ggcagcttcc acgcaggaag
     3001 gcaagtccca accattcaag gtcacgccgg ggccattcga tccagccaca tggctggaat
     3061 ggtcccgcca gtggcagggc actgaaggca acggccacgc ggccgcgtcc ggcattccgg
     3121 gcctggatgc gctggcaggc gtcaagatcg cgccggcgca gctgggtgat atccagcagc
     3181 gctacatgaa ggacttctca gcgctgtggc aggccatggc cgagggcaag gccgaggcca
     3241 ccggtccgct gcacgaccgg cgcttcgccg gcgacgcatg gcgcaccaac ctcccatatc
     3301 gcttcgctgc cgcgttctac ctgctcaatg cgcgcgcctt gaccgagctg gccgatgccg
     3361 tcgaggccga tgccaagacc cgccagcgca tccgcttcgc gatctcgcaa tgggtcgatg
     3421 cgatgtcgcc cgccaacttc cttgccacca atcccgaggc gcagcgcctg ctgatcgagt
     3481 cgggcggcga atcgctgcgt gccggcgtgc gcaacatgat ggaagacctg acacgcggca
     3541 agatctcgca gaccgacgag agcgcgtttg aggtcggccg caatgtcgcg gtgaccgaag
     3601 gcgccgtggt cttcgagaac gagtacttcc agctgttgca gtacaagccg ctgaccgaca
     3661 aggtgcacgc gcgcccgctg ctgatggtgc cgccgtgcat caacaagtac tacatcctgg
     3721 acctgcagcc ggagagctcg ctggtgcgcc atgtggtgga gcagggacat acggtgtttc
     3781 tggtgtcgtg gcgcaatccg gacgccagca tggccggcag cacctgggac gactacatcg
     3841 agcacgcggc catccgcgcc atcgaagtcg cgcgcgacat cagcggccag gacaagatca
     3901 acgtgctcgg cttctgcgtg ggcggcacca ttgtctcgac cgcgctggcg gtgctggccg
     3961 cgcgcggcga gcacccggcc gccagcgtca cgctgctgac cacgctgctg gactttgccg
     4021 acacgggcat cctcgacgtc tttgtcgacg agggccatgt gcagttgcgc gaggccacgc
     4081 tgggcggcgg cgccggcgcg ccgtgcgcgc tgctgcgcgg ccttgagctg gccaatacct
     4141 tctcgttctt gcgcccgaac gacctggtgt ggaactacgt ggtcgacaac tacctgaagg
     4201 gcaacacgcc ggtgccgttc gacctgctgt tctggaacgg cgacgccacc aacctgccgg
     4261 ggccgtggta ctgctggtac ctgcgccaca cctacctgca gaacgagctc aaggtaccgg
     4321 gcaagctgac cgtgtgcggc gtgccggtgg acctggccag catcgacgtg ccgacctata
     4381 tctacggctc gcgcgaagac catatcgtgc cgtggaccgc ggcctatgcc tcgaccgcgc
     4441 tgctggcgaa caagctgcgc ttcgtgctgg gtgcgtcggg ccatatcgcc ggtgtgatca
     4501 acccgccggc caagaacaag cgcagccact ggactaacga tgcgctgccg gagtcgccgc
     4561 agcaatggct ggccggcgcc atcgagcatc acggcagctg gtggccggac tggaccgcat
     4621 ggctggccgg gcaggccggc gcgaaacgcg ccgcgcccgc caactatggc aatgcgcgct
     4681 atcgcgcaat cgaacccgcg cctgggcgat acgtcaaagc caaggcatga cgcttgcatg
     4741 agtgccggcg tgcgtcatgc acggcgccgg caggcctgca ggttccctcc cgtttccatt
     4801 gaaaggacta cacaatgact gacgttgtca tcgtatccgc cgcccgcacc gcggtcggca
     4861 agtttggcgg ctcgctggcc aagatcccgg caccggaact gggtgccgtg gtcatcaagg
     4921 ccgcgctgga gcgcgccggc gtcaagccgg agcaggtgag cgaagtcatc atgggccagg
     4981 tgctgaccgc cggttcgggc cagaaccccg cacgccaggc cgcgatcaag gccggcctgc
     5041 cggcgatggt gccggccatg accatcaaca aggtgtgcgg ctcgggcctg aaggccgtga
     5101 tgctggccgc caacgcgatc atggcgggcg acgccgagat cgtggtggcc ggcggccagg
     5161 aaaacatgag cgccgccccg cacgtgctgc cgggctcgcg cgatggtttc cgcatgggcg
     5221 atgccaagct ggtcgacacc atgatcgtcg acggcctgtg ggacgtgtac aaccagtacc
     5281 acatgggcat caccgccgag aacgtggcca aggaatacgg catcacacgc gaggcgcagg
     5341 atgagttcgc cgtcggctcg cagaacaagg ccgaagccgc gcagaaggcc ggcaagtttg
     5401 acgaagagat cgtcccggtg ctgatcccgc agcgcaaggg cgacccggtg gccttcaaga
     5461 ccgacgagtt cgtgcgccag ggcgccacgc tggacagcat gtccggcctc aagcccgcct
     5521 tcgacaaggc cggcacggtg accgcggcca acgcctcggg cctgaacgac ggcgccgccg
     5581 cggtggtggt gatgtcggcg gccaaggcca aggaactggg cctgaccccg ctggccacga
     5641 tcaagagcta tgccaacgcc ggtgtcgatc ccaaggtgat gggcatgggc ccggtgccgg
     5701 cctccaagcg cgccctgtcg cgcgccgagt ggaccccgca agacctggac ctgatggaga
     5761 tcaacgaggc ctttgccgcg caggcgctgg cggtgcacca gcagatgggc tgggacacct
     5821 ccaaggtcaa tgtgaacggc ggcgccatcg ccatcggcca cccgatcggc gcgtcgggct
     5881 gccgtatcct ggtgacgctg ctgcacgaga tgaagcgccg tgacgcgaag aagggcctgg
     5941 cctcgctgtg catcggcggc ggcatgggcg tggcgctggc agtcgagcgc aaataaggaa
     6001 ggggttttcc ggggccgcgc gcggttggcg cggacccggc gacgataacg aagccaatca
     6061 aggagtggac atgactcagc gcattgcgta tgtgaccggc ggcatgggtg gtatcggaac
     6121 cgccatttgc cagcggctgg ccaaggatgg ctttcgtgtg gtggccggtt gcggccccaa
     6181 ctcgccgcgc cgcgaaaagt ggctggagca gcagaaggcc ctgggcttcg atttcattgc
     6241 ctcggaaggc aatgtggctg actgggactc gaccaagacc gcattcgaca aggtcaagtc
     6301 cgaggtcggc gaggttgatg tgctgatcaa caacgccggt atcacccgcg acgtggtgtt
     6361 ccgcaagatg acccgcgccg actgggatgc ggtgatcgac accaacctga cctcgctgtt
     6421 caacgtcacc aagcaggtga tcgacggcat ggccgaccgt ggctggggcc gcatcgtcaa
     6481 catctcgtcg gtgaacgggc agaagggcca gttcggccag accaactact ccaccgccaa
     6541 ggccggcctg catggcttca ccatggcact ggcgcaggaa gtggcgacca agggcgtgac
     6601 cgtcaacacg gtctctccgg gctatatcgc caccgacatg gtcaaggcga tccgccagga
     6661 cgtgctcgac aagatcgtcg cgacgatccc ggtcaagcgc ctgggcctgc cggaagagat
     6721 cgcctcgatc tgcgcctggt tgtcgtcgga ggagtccggt ttctcgaccg gcgccgactt
     6781 ctcgctcaac ggcggcctgc atatgggctg atgggctgaa gggtttaagc gaagacgaac
     6841 aataaggcct cccaaatcgg ggggcctttt ttattgataa caaaaaggca acactatgac
     6901 atcggaaaac ccgttactgg cgctgcgaga gaaaatcagc gcgctggatg aaaaattatt
     6961 agcgttactg gcagagaggc gcgaattcaa acagaaaggc catgggctgg cctcaggcat
     7021 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
     7081 cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat ttgctttcga
     7141 atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg
     7201 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     7261 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     7321 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     7381 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     7441 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     7501 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     7561 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     7621 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
     7681 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     7741 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     7801 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     7861 atatccagtg atttttttct ccattttagc ttccttagct cctgaa
//
