LOCUS       sequence-238544-        8345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a designed construct of the condensing part fused to the 
            ACP domain of murine type I fatty acid synthase (FASN) in 
            Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  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 8345)
  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: pAR127_StrepI_m(KS_MAT)_mouseL_ACP_H8_pET22b
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..79
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     163..182
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        163..181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    182..206
                     /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             221..243
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(254..259,260..283,284..319,320..343)
                     /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             3233..3256
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3309..3327)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3323..3370
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3407..3862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3494..3513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3704..3725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3888..3992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3993..4061,4062..4853)
                     /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(4211..4230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5024..5612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5513..5532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5766..5783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5798..5937
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5863..5882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6023..6045
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6039..6230)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7039..8121)
                     /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     8083..8102
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8122..8199)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
       61 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
      121 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
      181 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
      241 agatatacat atgagcgctt ggagccatcc acaatttgag aagggtggag gttctggcgg
      301 tggatcggga ggttcagcgt ggagccaccc gcagttcgaa aaaggcgccg gatccgagga
      361 ggtggtgata gccggtatgt cggggaagtt gcccgagtca gagaacctac aggagttctg
      421 ggccaacctc attggtggtg tggacatggt cacagatgat gacaggagat ggaaggctgg
      481 gctctatgga ttacccaagc ggtctggaaa gctgaaggat ctctccaagt tcgacgcctc
      541 cttttttggg gtccacccca agcaggcaca cacaatggac ccccagcttc ggctgctgtt
      601 ggaagtcagc tatgaagcaa ttgtggatgg aggtatcaac ccagcctcac tccgaggaac
      661 gaacactggc gtctgggtgg gtgtgagtgg ttcagaggca tccgaggccc ttagcagaga
      721 tcccgagacg cttctgggct acagcatggt gggctgccag cgtgcaatga tggccaaccg
      781 gctctctttc ttcttcgact tcaaaggacc aagcattgcc ctggacacag cctgctcctc
      841 cagcttgctg gcactacaga atgcctacca ggccatccgt agtggggaat gccccgcggc
      901 ccttgtgggt gggatcaacc tgctcctgaa gccgaacacc tctgtgcagt tcatgaagct
      961 gggcatgctc agcccggacg gcacctgcag atcctttgat gattcaggga gtggatattg
     1021 tcgctctgag gctgttgtag cagttctgct gactaagaag tccctggctc ggcgggtcta
     1081 tgccacgatt ctgaatgccg gcaccaatac agatggcagc aaggagcaag gtgtaacatt
     1141 cccctctgga gaagtccaag aacaactcat ctgctctctg tatcagccag ctggtctggc
     1201 cccggagtcg cttgagtata ttgaagccca tggcacgggc accaaggtgg gtgaccccca
     1261 ggaactgaat ggcattactc ggtccctgtg cgccttccgc caggcccctc tgttaattgg
     1321 ctccaccaaa tccaacatgg gacaccctga gcctgcctct gggcttgcag ccctgaccaa
     1381 ggtgctgtta tccctggagc atggggtctg ggcccctaac ctgcacttcc acaaccccaa
     1441 ccctgagatc ccagcacttc ttgatgggcg gctgcaggtg gtcgataggc ccctgcctgt
     1501 tcgtggtggc aacgtgggca tcaactcatt tggcttcgga ggctccaatg ttcatgtcat
     1561 cctccagccc aacacacggc aggcccctgc gcccactgca cacgctgccc ttccccattt
     1621 gctgcacgcc agtggacgca ccttagaggc agtgcaggac ctgctggaac agggccgcca
     1681 gcacagccag gacctggcct ttgtgagcat gctcaatgac attgcggcaa cccctacagc
     1741 agccatgccc ttcaggggtt acactgtgct aggtgttgag ggccgtgtcc aagaagtgca
     1801 gcaagtgtcc accaacaagc gcccactctg gttcatctgc tcagggatgg gcacgcagtg
     1861 gcgcgggatg gggctgagcc tcatgcgcct ggacagcttc cgtgagtcta tcctgcgctc
     1921 cgatgaggct gtgaagccgt tgggagtgaa agtgtcagat ctgctgttga gcacagatga
     1981 gcgcaccttt gatgacatcg tgcatgcctt tgtgagcctc actgccatcc agattgccct
     2041 catcgaccta ctgacttctg tgggactgaa acctgacggc atcattgggc actccttggg
     2101 agaggttgcc tgtggctatg cagatggctg tctctcccag agagaggctg tgcttgcagc
     2161 ttactggcga ggccagtgca tcaaagatgc ccacctcccg cctggatcca tggcagctgt
     2221 tggtttgtcc tgggaggaat gtaaacagcg ctgccccgct ggcgtggtgc ctgcctgcca
     2281 caactctgag gacaccgtga ccatctctgg acctcaggct gcagtgaatg aatttgtgga
     2341 gcagctaaag caagaaggtg tgtttgccaa ggaggtacga acaggaggcc tggctttcca
     2401 ctcctacttc atggaaggaa ttgcccccac attgctgcag gctctcaaga aggtgatccg
     2461 ggaaccacgg ccgcgctcgg ctcgatggct cagcacctct atccctgagg cccagtggca
     2521 gagcagcctg gcccgcacat cttctgccga gtacaatgtc aacaacctgg tgagccctgt
     2581 gctcttccag gaagcactgt ggcacatccc tgagcatgcc gtggtgctgg agattgcgcc
     2641 ccacgcactg ttgcaggctg tcctgaagcg aggcgtgaag tccagctgca ccatcattcc
     2701 cttgatgaag agggatcata aagataactt ggagttcttt ctcaccaacc ttggcaaggt
     2761 gcacctcaca ggcatcaatg tcaaccctaa cgccttgttc ccacctgtgg agttcccggc
     2821 tccccgaggg actcctctca tctcccctca catcaagtgg gaccacagtc agacttggga
     2881 tgtcccggtt gctgaggact tcccaaacgg ctccagctcc tcctctgcta cagtctacag
     2941 catcgacgcc agtgcagaga agaaagctgt ggcccatggg gacggggaca cccagaggga
     3001 tctggtgaaa gctgtagcac acatcctagg catccgagac ctcgcaggta ttaacctgga
     3061 cagcacgctg gcagacctcg gcctggactc gctcatgggt gtggaagttc gtcagatcct
     3121 ggaacgagaa cacgatctgg tgctgcccat gcgtgaggtg cggcagctca cgctgcggaa
     3181 acttcaggaa atgtcctcca agactgactc ggctactgac acgacactcg agcatcatca
     3241 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     3301 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     3361 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     3421 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3481 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3541 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3601 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3661 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3721 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3781 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3841 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3901 tttgtttatt tttctaaata cattcaaata tgtattcgct catgagacaa taaccctgat
     3961 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4021 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4081 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4141 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4201 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4261 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4321 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4381 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4441 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4501 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     4561 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4621 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4681 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4741 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4801 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4861 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4921 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4981 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5041 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5101 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5161 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5221 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5281 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5341 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5401 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5461 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5521 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5581 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5641 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5701 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5761 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     5821 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     5881 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     5941 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6001 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     6061 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     6121 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     6181 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     6241 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     6301 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     6361 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     6421 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     6481 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     6541 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     6601 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     6661 gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg
     6721 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     6781 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     6841 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     6901 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     6961 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     7021 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7081 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     7141 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     7201 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     7261 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     7321 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     7381 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     7441 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     7501 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     7561 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     7621 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     7681 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     7741 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     7801 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     7861 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     7921 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     7981 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     8041 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     8101 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     8161 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     8221 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     8281 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     8341 ccacc
//
