LOCUS       sequence-259536-        6641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses S. thermophilus DltB in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6641)
  AUTHORS   Ma D, Wang Z, Merrikh CN, Lang KS, Lu P, Li X, Merrikh H, Rao Z, Xu 
            W
  TITLE     Crystal structure of a membrane-bound O-acyltransferase.
  JOURNAL   Nature. 2018 Oct;562(7726):286-290. doi: 10.1038/s41586-018-0568-2. 
            Epub 2018 Oct 3.
  PUBMED    30283133
REFERENCE   2  (bases 1 to 6641)
  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"; doi: 
            "10.1038/s41586-018-0568-2"; journalName: "Nature"; date: "2018-10";
            volume: "562"; issue: "7726"; pages: "286-290"
COMMENT     Sequence Label: pET21b - S. thermophilus DltB
FEATURES             Location/Qualifiers
     source          1..6641
                     /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             1321..1344
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1354..1371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1424..1442)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1438..1485
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1522..1977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1609..1628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1819..1840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2003..2107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2108..2176,2177..2968)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2326..2345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3139..3727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3628..3647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3881..3898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3913..4052
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3978..3997)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4138..4160
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4154..4345)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5154..6236)
                     /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     6198..6217
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6237..6314)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6520..6539
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6623..6641,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6623..6641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tgattgactt cctgaaacaa ctgccgcatc tggagccgta tggtaacccg
      121 ttctacttta tttatctggg tatcgcgctg ctgccgattt tcattggcct gttctttaag
      181 aaacgttttg cgatttacga gtgcctggtt agcatcacct tcattgttct ggcgctgacc
      241 ggcacccatg cgagccagat tctggcgctg ctgttttata ttgtgtggca aatcatttgg
      301 gtttacagct ataaacgtta ccgtagccag cgtgataaca agtgggtgtt ttatctgcac
      361 agcttcctgg tggttctgcc gctgattctg gtgaaagttg agccgaccat taacggcacc
      421 caaagcctgc tgaacttcct gggtatcagc tacctgacct ttcgtgcggt gggtatgatt
      481 atcgaaatgc gtgacggcgt tctgaaagag ttcaccctgg gcgagttcct gcgttttatg
      541 ctgttcatgc cgacctttac cagcggcccg attgaccgtt ttaagcgttt caacgaagat
      601 tatcagagca ttccgaaccg tgacgagctg ctgaacatgc tggaacaagc ggtgaaatac
      661 atcatgctgg gctttctgta taagttcgtt ctggcgcaga ttttcggtag catgctgctg
      721 ccgccgctga aagcgcaggc gctgagccaa ggtggcatct tcaacctgcc gaccctgggt
      781 gtgatgtacg tttatggctt tgacctgttc tttgatttcg cgggctacag catgtttgcg
      841 ctggcggtta gcaacctgat gggtatcaaa agcccgatta actttgacaa gccgttcatc
      901 agccgtgata tgaaggagtt ctggaaccgt tggcacatga gcctgagctt ctggtttcgt
      961 gatttcgtgt ttatgcgtct ggtgattgtt ctgatgcgta acaaagtttt caagaaccgt
     1021 aacaccacca gcaacgtggc gtacatcatt aacatgatgg ttatgggttt ctggcacggc
     1081 atcacctggt actatatcgc gtatggtatt tttcacggta tcggcctggt gattaacgat
     1141 gcgtggctgc gtaagaaaaa gaccattaac aaagaccgta aaaaggcggg cctgaagccg
     1201 ctgccggaaa acaaatggac caaggcgctg ggtatcttta ttaccttcaa caccgttatg
     1261 ctgagcttcc tgatttttag cggcttcctg aatgacctgt ggtttaccaa gaaactcgag
     1321 catcaccatc accatcacca tcactgagtc gagcaccacc accaccacca ctgagatccg
     1381 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1441 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1501 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1561 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1621 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1681 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1741 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1801 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1861 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1921 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     1981 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2101 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2161 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2221 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2281 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2341 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2401 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2461 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2521 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2581 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2641 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2701 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2761 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2821 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2881 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2941 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3001 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3061 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3241 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3361 cttgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3781 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3841 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3901 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3961 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     4021 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     4081 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4141 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4201 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4261 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4321 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     4381 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     4441 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     4501 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     4561 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     4621 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     4681 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     4741 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     4801 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     4861 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     4921 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     4981 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     5041 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     5101 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     5161 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5221 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     5281 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     5341 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     5401 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     5461 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     5521 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     5581 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     5641 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     5701 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     5761 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     5821 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     5881 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     5941 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     6001 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     6061 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     6121 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     6181 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     6241 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     6301 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     6361 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     6421 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     6481 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     6541 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     6601 cgagatctcg atcccgcgaa attaatacga ctcactatag g
//
