LOCUS       sequence-222947-        9616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ads under Pgrac promoter (inducible by IPTG) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9616)
  AUTHORS   Zhou K, Zou R, Zhang C, Stephanopoulos G, Too HP
  TITLE     Optimization of amorphadiene synthesis in Bacillus subtilis via 
            transcriptional, translational, and media modulation.
  JOURNAL   Biotechnol Bioeng. 2013 Mar 8. doi: 10.1002/bit.24900.
  PUBMED    23483530
REFERENCE   2  (bases 1 to 9616)
  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: "Biotechnol 
            Bioeng. 2013 Mar 8. doi: 10.1002/bit.24900."
COMMENT     Sequence Label: pHT-ads
FEATURES             Location/Qualifiers
     source          1..9616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1224..1246
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1238..1255
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1239..1255
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1298..2380)
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2342..2361
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    2764..2788
                     /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)."
     CDS             4448..4465
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        7693..7797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7798..7866,7867..8658)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8016..8035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8829..9417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9318..9337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9571..9588
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttaagttatt ggtatgactg gttttaagcg caaaaaaagt tgctttttcg tacctattaa
       61 tgtatcgttt tagaaaaccg actgtaaaaa gtacagtcgg cattatctca tattataaaa
      121 gccagtcatt aggcctatct gacaattcct gaatagagtt cataaacaat cctgcatgat
      181 aaccatcaca aacagaatga tgtacctgta aagatagcgg taaatatatt gaattacctt
      241 tattaatgaa ttttcctgct gtaataatgg gtagaaggta attactatta ttattgatat
      301 ttaagttaaa cccagtaaat gaagtccatg gaataataga aagagaaaaa gcattttcag
      361 gtataggtgt tttgggaaac aatttccccg aaccattata tttctctaca tcagaaaggt
      421 ataaatcata aaactctttg aagtcattct ttacaggagt ccaaatacca gagaatgttt
      481 tagatacacc atcaaaaatt gtataaagtg gctctaactt atcccaataa cctaactctc
      541 cgtcgctatt gtaaccagtt ctaaaagctg tatttgagtt tatcaccctt gtcactaaga
      601 aaataaatgc agggtaaaat ttatatcctt cttgttttat gtttcggtat aaaacactaa
      661 tatcaatttc tgtggttata ctaaaagtcg tttgttggtt caaataatga ttaaatatct
      721 cttttctctt ccaattgtct aaatcaattt tattaaagtt catttgatat gcctcctaaa
      781 tttttatcta aagtgaattt aggaggctta cttgtctgct ttcttcatta gaatcaatcc
      841 ttttttaaaa gtcaatatta ctgtaacata aatatatatt ttaaaaatat cccactttat
      901 ccaattttcg tttgttgaac taatgggtgc tttagttgaa gaataaaaga ccacattaaa
      961 aaatgtggtc ttttgtgttt ttttaaagga tttgagcgta gcgaaaaatc cttttctttc
     1021 ttatcttgat aataagggta actattgccg atcgtccatt ccgacagcat cgccagtcac
     1081 tatggcgtgc tgctagcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1141 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     1201 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     1261 cgagctcagg ccttaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1321 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1381 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     1441 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     1501 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     1561 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     1621 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     1681 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     1741 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     1801 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     1861 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     1921 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     1981 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     2041 gattgtgcac cgccgtttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     2101 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     2161 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     2221 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     2281 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     2341 cggcatactc tgcgacatcg tataacgtta ctggtttcat caaaatcgtc tccctccgtt
     2401 tgaatatttg attgatcgta accagatgaa gcactctttc cactatccct acagtgttat
     2461 ggcttgaaca atcacgaaac aataattggt acgtacgatc tttcagccga ctcaaacatc
     2521 aaatcttaca aatgtagtct ttgaaagtat tacatatgta agatttaaat gcaaccgttt
     2581 tttcggaagg aaatgatgac ctcgtttcca ccggaattag cttggtacca gctattgtaa
     2641 cataatcggt acgggggtga aaaagctaac ggaaaaggga gcggaaaaga atgatgtaag
     2701 cgtgaaaaat tttttatctt atcacttgaa attggaaggg agattcttta ttataagaat
     2761 tgtggaattg tgagcggata acaattccca attaaaggag gaaggatcca tgtctctgac
     2821 tgaggaaaaa ccaatccgtc cgatcgcaaa ctttccgccg agcatctggg gtgatcagtt
     2881 cctgatctac cagaagcagg tcgagcaagg cgtagaacag atcgtgaacg acctgaaaaa
     2941 ggaggtacgt cagctgctga aagaggctct ggacatcccg atgaaacatg cgaacctgct
     3001 gaagctgatt gatgagattc agcgcctggg tatcccatac cacttcgaac gtgaaattga
     3061 tcacgcgctg cagtgtattt atgaaacgta tggtgataac tggaacggtg accgtagctc
     3121 cctgtggttc cgtctgatgc gtaagcaggg ttattacgtg acctgcgacg tcttcaacaa
     3181 ctacaaagac aagaacggcg cgtttaaaca gagcctggcg aatgacgttg aaggcctgct
     3241 ggagctgtat gaagcaacct ctatgcgtgt tccgggcgaa atcatcctgg aagatgctct
     3301 gggtttcacc cgttctcgcc tgtctatcat gaccaaggac gcattttcca ctaacccggc
     3361 cctgttcacc gaaatccagc gtgcgctgaa acagcctctg tggaagcgtc tgccgcgtat
     3421 cgaggcggca cagtacatcc cgttctatca gcaacaggat tcccacaaca aaaccctgct
     3481 gaaactggcg aaactggaat ttaacctgct gcaatccctg cacaaagaag aactgtctca
     3541 tgtttgcaaa tggtggaaag ccttcgacat caagaaaaac gcgccgtgcc tgcgcgaccg
     3601 tatcgtagaa tgctatttct ggggcctggg ctccggttat gagcctcaat actctcgtgc
     3661 tcgcgtattc ttcaccaaag cggtagctgt gatcaccctg atcgacgata cctacgatgc
     3721 ttacggtacg tacgaagaac tgaagatttt taccgaggct gttgaacgct ggtctatcac
     3781 ttgcctggat accctgccgg aatacatgaa accgatctat aaactgttca tggatactta
     3841 taccgaaatg gaagagttcc tggcaaaaga aggtcgcact gatctgttca actgtggcaa
     3901 ggagtttgtg aaagaattcg tccgcaacct gatggtggag gcgaagtggg caaacgaggg
     3961 tcacatcccg accaccgaag aacacgaccc ggtagttatc atcaccggtg gcgcaaacct
     4021 gctgactacc acttgctacc tgggtatgtc cgacattttt acgaaagaaa gcgttgaatg
     4081 ggcagtgtct gcgcctccgc tgttccgtta ctctggtatc ctgggccgtc gcctgaacga
     4141 tctgatgacc cataaagccg agcaggaacg taaacactcc agcagctctc tggaatctta
     4201 catgaaagaa tacaacgtta acgaagagta tgcgcagacc ctgatctaca aagaggttga
     4261 ggatgtatgg aaagacatca accgtgagta cctgactact aaaaacatcc cacgcccgct
     4321 gctgatggca gtgatttatc tgtgccagtt tctggaagtg cagtatgcgg gcaaagataa
     4381 cttcacccgt atgggtgacg aatacaaaca cctgatcaag tccctgctgg tttacccaat
     4441 gtccatccat catcatcacc atcactgatc tagagtcgac gtccccgggg cagcccgcct
     4501 aatgagcggg cttttttcac gtcacgcgtc catggagatc tttgtctgca actgaaaagt
     4561 ttatacctta gcctggaaca aatggttgaa acatacgagg ctaatatcgg cttattagga
     4621 atagtccctg tactaataaa atcaggtgga tcagttgatc agtatatttt ggacgaagct
     4681 cggaaagaat ttggagatga cttgcttaat tccacaatta aattaaggga aagaataaag
     4741 cgatttgatg ttcaaggaat cacggaagaa gatactcatg ataaagaagc tctaaaacta
     4801 ttcaataacc ttacaatgga attgatcgaa agggtggaag gttaatggta cgaaaattag
     4861 gggatctacc tagaaagcca caaggcgata ggtcaagctt aaagaaccct tacatggatc
     4921 ttacagattc tgaaagtaaa gaaacaacag aggttaaaca aacagaacca aaaagaaaaa
     4981 aagcattgtt gaaaacaatg aaagttgatg tttcaatcca taataagatt aaatcgctgc
     5041 acgaaattct ggcagcatcc gaagggaatt catattactt agaggatact attgagagag
     5101 ctattgataa gatggttgag acattacctg agagccaaaa aactttttat gaatatgaat
     5161 taaaaaaaag aaccaacaaa ggctgagaca gactccaaac gagtctgttt ttttaaaaaa
     5221 aatattagga gcattgaata tatattagag aattaagaaa gacatgggaa taaaaatatt
     5281 ttaaatccag taaaaatatg ataagattat ttcagaatat gaagaactct gtttgttttt
     5341 gatgaaaaaa caaacaaaaa aaatccacct aacggaatct caatttaact aacagcggcc
     5401 aaactgagaa gttaaatttg agaaggggaa aaggcggatt tatacttgta tttaactatc
     5461 tccattttaa cattttatta aaccccatac aagtgaaaat cctcttttac actgttcctt
     5521 taggtgatcg cggagggaca ttatgagtga agtaaaccta aaaggaaata cagatgaatt
     5581 agtgtattat cgacagcaaa ccactggaaa taaaatcgcc aggaagagaa tcaaaaaagg
     5641 gaaagaagaa gtttattatg ttgctgaaac ggaagagaag atatggacag aagagcaaat
     5701 aaaaaacttt tctttagaca aatttggtac gcatatacct tacatagaag gtcattatac
     5761 aatcttaaat aattacttct ttgatttttg gggctatttt ttaggtgctg aaggaattgc
     5821 gctctatgct cacctaactc gttatgcata cggcagcaaa gacttttgct ttcctagtct
     5881 acaaacaatc gctaaaaaaa tggacaagac tcctgttaca gttagaggct acttgaaact
     5941 gcttgaaagg tacggtttta tttggaaggt aaacgtccgt aataaaacca aggataacac
     6001 agaggaatcc ccgattttta agattagacg taaggttcct ttgctttcag aagaactttt
     6061 aaatggaaac cctaatattg aaattccaga tgacgaggaa gcacatgtaa agaaggcttt
     6121 aaaaaaggaa aaagagggtc ttccaaaggt tttgaaaaaa gagcacgatg aatttgttaa
     6181 aaaaatgatg gatgagtcag aaacaattaa tattccagag gccttacaat atgacacaat
     6241 gtatgaagat atactcagta aaggagaaat tcgaaaagaa atcaaaaaac aaatacctaa
     6301 tcctacaaca tcttttgaga gtatatcaat gacaactgaa gaggaaaaag tcgacagtac
     6361 tttaaaaagc gaaatgcaaa atcgtgtctc taagccttct tttgatacct ggtttaaaaa
     6421 cactaagatc aaaattgaaa ataaaaattg tttattactt gtaccgagtg aatttgcatt
     6481 tgaatggatt aagaaaagat atttagaaac aattaaaaca gtccttgaag aagctggata
     6541 tgttttcgaa aaaatcgaac taagaaaagt gcaataaact gctgaagtat ttcagcagtt
     6601 ttttttattt agaaatagtg aaaaaaatat aatcagggag gtatcaatat ttaatgagta
     6661 ctgatttaaa tttatttaga ctggaattaa taattaacac gtagactaat taaaatttaa
     6721 tgagggataa agaggataca aaaatattaa tttcaatccc tattaaattt taacaagggg
     6781 gggattaaaa tttaattaga ggtttatcca caagaaaaga ccctaataaa atttttacta
     6841 gggttataac actgattaat ttcttaatgg gggagggatt aaaatttaat gacaaagaaa
     6901 acaatctttt aagaaaagct tttaaaagat aataataaaa agagctttgc gattaagcaa
     6961 aactctttac tttttcattg acattatcaa attcatcgat ttcaaattgt tgttgtatca
     7021 taaagttaat tctgttttgc acaacctttt caggaatata aaacacatct gaggcttgtt
     7081 ttataaactc agggtcgcta aagtcaatgt aacgtagcat atgatatggt atagcttcca
     7141 cccaagttag cctttctgct tcttctgaat gtttttcata tacttccatg ggtatctcta
     7201 aatgattttc ctcatgtagc aaggtatgag caaaaagttt atggaattga tagttcctct
     7261 ctttttcttc aactttttta tctaaaacaa acactttaac atctgagtca atgtaagcat
     7321 aagatgtttt tccagtcata atttcaatcc caaatctttt agacagaaat tctggacgta
     7381 aatcttttgg tgaaagaatt tttttatgta gcaatatatc cgatacagca ccttctaaaa
     7441 gcgttggtga atagggcatt ttacctatct cctctcattt tgtggaataa aaatagtcat
     7501 attcgtccat ctacctatcc tattatcgaa cagttgaact ttttaatcaa ggatcagtcc
     7561 tttttttcat tattcttaaa ctgtgctctt aactttaaca actcgatttg tttttccaga
     7621 tctcgagggt aactagcctc gccgatcccg caagaggccc ggcagtcagg tggcactttt
     7681 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     7741 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7801 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7861 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7921 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7981 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     8041 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     8101 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8161 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8221 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8281 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8341 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8401 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8461 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     8521 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     8581 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     8641 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     8701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     8761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     8821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8941 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     9001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9421 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     9481 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     9541 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     9601 cgcccaatac gcatgc
//
