LOCUS       sequence-238542-        8042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the condensing part of murine type I fatty acid synthase 
            (FASN) consisting of the KS and MAT domain in Escherichia coli. 
            N-terminal Twin-Strep tag; C-terminal H8-tag; in pET22b vector. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8042)
  AUTHORS   Rittner A, Paithankar KS, Drexler DJ, Himmler A, Grininger M
  TITLE     Probing the modularity of megasynthases by rational engineering of a
            fatty acid synthase Type I.
  JOURNAL   Protein Sci. 2019 Feb;28(2):414-428. doi: 10.1002/pro.3550. Epub 
            2018 Dec 20.
  PUBMED    30394635
REFERENCE   2  (bases 1 to 8042)
  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: "10.1002/pro.3550"; 
            journalName: "Protein Sci"; date: "2019-02"; volume: "28"; issue: 
            "2"; pages: "414-428"
COMMENT     Sequence Label: pAR69_STRI_m(KS_MAT)_H8_pET22b
FEATURES             Location/Qualifiers
     source          1..8042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..55
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     139..158
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        139..157
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    158..182
                     /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             197..219
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(230..235,236..259,260..295,296..319)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             2906..2929
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2982..3000)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2996..3043
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3080..3535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3167..3186)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3377..3398
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3561..3665
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3666..3734,3735..4526)
                     /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(3884..3903)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4697..5285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5186..5205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5439..5456
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5471..5610
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5536..5555)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5696..5718
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5712..5903)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6712..7794)
                     /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     7756..7775
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7795..7872)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
       61 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
      121 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
      181 cccctctaga aataattttg tttaacttta agaaggagat atacatatga gcgcttggag
      241 ccatccacaa tttgagaagg gtggaggttc tggcggtgga tcgggaggtt cagcgtggag
      301 ccacccgcag ttcgaaaaag gcgccggatc cgaggaggtg gtgatagccg gtatgtcggg
      361 gaagttgccc gagtcagaga acctacagga gttctgggcc aacctcattg gtggtgtgga
      421 catggtcaca gatgatgaca ggagatggaa ggctgggctc tatggattac ccaagcggtc
      481 tggaaagctg aaggatctct ccaagttcga cgcctccttt tttggggtcc accccaagca
      541 ggcacacaca atggaccccc agcttcggct gctgttggaa gtcagctatg aagcaattgt
      601 ggatggaggt atcaacccag cctcactccg aggaacgaac actggcgtct gggtgggtgt
      661 gagtggttca gaggcatccg aggcccttag cagagatccc gagacgcttc tgggctacag
      721 catggtgggc tgccagcgtg caatgatggc caaccggctc tctttcttct tcgacttcaa
      781 aggaccaagc attgccctgg acacagcctg ctcctccagc ttgctggcac tacagaatgc
      841 ctaccaggcc atccgtagtg gggaatgccc cgcggccctt gtgggtggga tcaacctgct
      901 cctgaagccg aacacctctg tgcagttcat gaagctgggc atgctcagcc cggacggcac
      961 ctgcagatcc tttgatgatt cagggagtgg atattgtcgc tctgaggctg ttgtagcagt
     1021 tctgctgact aagaagtccc tggctcggcg ggtctatgcc acgattctga atgccggcac
     1081 caatacagat ggcagcaagg agcaaggtgt aacattcccc tctggagaag tccaagaaca
     1141 actcatctgc tctctgtatc agccagctgg tctggccccg gagtcgcttg agtatattga
     1201 agcccatggc acgggcacca aggtgggtga cccccaggaa ctgaatggca ttactcggtc
     1261 cctgtgcgcc ttccgccagg cccctctgtt aattggctcc accaaatcca acatgggaca
     1321 ccctgagcct gcctctgggc ttgcagccct gaccaaggtg ctgttatccc tggagcatgg
     1381 ggtctgggcc cctaacctgc acttccacaa ccccaaccct gagatcccag cacttcttga
     1441 tgggcggctg caggtggtcg ataggcccct gcctgttcgt ggtggcaacg tgggcatcaa
     1501 ctcatttggc ttcggaggct ccaatgttca tgtcatcctc cagcccaaca cacggcaggc
     1561 ccctgcgccc actgcacacg ctgcccttcc ccatttgctg cacgccagtg gacgcacctt
     1621 agaggcagtg caggacctgc tggaacaggg ccgccagcac agccaggacc tggcctttgt
     1681 gagcatgctc aatgacattg cggcaacccc tacagcagcc atgcccttca ggggttacac
     1741 tgtgctaggt gttgagggcc gtgtccaaga agtgcagcaa gtgtccacca acaagcgccc
     1801 actctggttc atctgctcag ggatgggcac gcagtggcgc gggatggggc tgagcctcat
     1861 gcgcctggac agcttccgtg agtctatcct gcgctccgat gaggctgtga agccgttggg
     1921 agtgaaagtg tcagatctgc tgttgagcac agatgagcgc acctttgatg acatcgtgca
     1981 tgcctttgtg agcctcactg ccatccagat tgccctcatc gacctactga cttctgtggg
     2041 actgaaacct gacggcatca ttgggcactc cttgggagag gttgcctgtg gctatgcaga
     2101 tggctgtctc tcccagagag aggctgtgct tgcagcttac tggcgaggcc agtgcatcaa
     2161 agatgcccac ctcccgcctg gatccatggc agctgttggt ttgtcctggg aggaatgtaa
     2221 acagcgctgc cccgctggcg tggtgcctgc ctgccacaac tctgaggaca ccgtgaccat
     2281 ctctggacct caggctgcag tgaatgaatt tgtggagcag ctaaagcaag aaggtgtgtt
     2341 tgccaaggag gtacgaacag gaggcctggc tttccactcc tacttcatgg aaggaattgc
     2401 ccccacattg ctgcaggctc tcaagaaggt gatccgggaa ccacggccgc gctcggctcg
     2461 atggctcagc acctctatcc ctgaggccca gtggcagagc agcctggccc gcacatcttc
     2521 tgccgagtac aatgtcaaca acctggtgag ccctgtgctc ttccaggaag cactgtggca
     2581 catccctgag catgccgtgg tgctggagat tgcgccccac gcactgttgc aggctgtcct
     2641 gaagcgaggc gtgaagtcca gctgcaccat cattcccttg atgaagaggg atcataaaga
     2701 taacttggag ttctttctca ccaaccttgg caaggtgcac ctcacaggca tcaatgtcaa
     2761 ccctaacgcc ttgttcccac ctgtggagtt cccggctccc cgagggactc ctctcatctc
     2821 ccctcacatc aagtgggacc acagtcagac ttgggatgtc ccggttgctg aggacttccc
     2881 aaacggctca ggttccccct cagcccatca tcaccaccac caccaccact gagatccggc
     2941 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     3001 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     3061 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3121 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3181 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3241 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3301 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3361 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3421 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3481 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3541 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3601 atatgtattc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3661 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3721 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3781 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3841 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3901 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3961 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4021 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4081 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4141 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4201 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4261 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4321 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4381 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4441 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4501 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4561 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4621 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4681 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4741 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4801 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4861 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4921 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4981 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5041 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5101 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5161 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5221 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5281 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5341 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5401 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5461 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5521 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     5581 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     5641 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5701 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     5761 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     5821 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     5881 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     5941 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     6001 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     6061 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     6121 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     6181 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     6241 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     6301 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     6361 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     6421 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     6481 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     6541 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     6601 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     6661 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     6721 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6781 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     6841 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     6901 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     6961 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     7021 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     7081 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     7141 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     7201 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     7261 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     7321 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     7381 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     7441 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     7501 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     7561 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     7621 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     7681 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     7741 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     7801 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     7861 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     7921 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     7981 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     8041 tc
//
