LOCUS       sequence-391053-        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MerTK Kinase Domain (wild-type) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  AUTHORS   Bae SH, Kim JH, Park TH, Lee K, Lee BI, Jang H
  TITLE     BMS794833 inhibits macrophage efferocytosis by directly binding to 
            MERTK and inhibiting its activity.
  JOURNAL   Exp Mol Med. 2022 Sep;54(9):1450-1460. doi: 
            10.1038/s12276-022-00840-x. Epub 2022 Sep 2.
  PUBMED    36056187
REFERENCE   2  (bases 1 to 7226)
  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/s12276-022-00840-x"; journalName: "Exp Mol Med"; date: 
            "2022-09"; volume: "54"; issue: "9"; pages: "1450-1460"
COMMENT     Sequence Label: pETDuet-MerTK-PTPN1
FEATURES             Location/Qualifiers
     source          1..7226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        99..117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    118..142
                     /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             157..179
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             198..215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             225..242
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        1193..1211
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1212..1236
                     /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             2290..2334
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2372..2390)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2386..2433
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2470..2925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2557..2576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2767..2788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3040..3831,3832..3900))
                     /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     3663..3682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3901..3993)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4074..4662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4563..4582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4816..4833
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4848..4987
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4913..4932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5073..5095
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5089..5280)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5852..6934)
                     /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     6896..6915
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6935..7012)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(7218..7226,1..11)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
       61 ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta atacgactca ctatagggga
      121 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      181 ataccatggg cagcagccat catcatcatc atcacagcag cggcctggtt ccgcgtggta
      241 gtgaggaact acaaaataaa ctagaagatg ttgtgattga caggaatctt ctaattcttg
      301 gaaaaattct gggtgaagga gagtttgggt ctgtaatgga aggaaatctt aagcaggaag
      361 atgggacctc tctgaaagtg gcagtgaaga ccatgaagtt ggacaactct tcacagcggg
      421 agatcgagga gtttctcagt gaggcagcgt gcatgaaaga cttcagccac ccaaatgtca
      481 ttcgacttct aggtgtgtgt atagaaatga gctctcaagg catcccaaag cccatggtaa
      541 ttttaccctt catgaaatac ggggacctgc atacttactt actttattcc cgattggaga
      601 caggaccaaa gcatattcct ctgcagacac tattgaagtt catggtggat attgccctgg
      661 gaatggagta tctgagcaac aggaattttc ttcatcgaga tttagctgct cgaaactgca
      721 tgttgcgaga tgacatgact gtctgtgttg cggacttcgg cctctctaag aagatttaca
      781 gtggcgatta ttaccgccaa ggccgcattg ctaagatgcc tgttaaatgg atcgccatag
      841 aaagtcttgc agaccgagtc tacacaagta aaagtgatgt gtgggcattt ggcgtgacca
      901 tgtgggaaat agctacgcgg ggaatgactc cctatcctgg ggtccagaac catgagatgt
      961 atgactatct tctccatggc cacaggttga agcagcccga agactgcctg gatgaactgt
     1021 atgaaataat gtactcttgc tggagaaccg atcccttaga ccgccccacc ttttcagtat
     1081 tgaggctgca gctagaaaaa ctcttagaaa gtttgcctga cgtttgagcg gccgcataat
     1141 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
     1201 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
     1261 taagaaggag atatacatat ggagatggaa aaggagttcg agcagatcga caagtccggg
     1321 agctgggcgg ccatttacca ggatatccga catgaagcca gtgacttccc atgtagagtg
     1381 gccaagcttc ctaagaacaa aaaccgaaat aggtacagag acgtcagtcc ctttgaccat
     1441 agtcggatta aactacatca agaagataat gactatatca acgctagttt gataaaaatg
     1501 gaagaagccc aaaggagtta cattcttacc cagggccctt tgcctaacac atgcggtcac
     1561 ttttgggaga tggtgtggga gcagaaaagc aggggtgtcg tcatgctcaa cagagtgatg
     1621 gagaaaggtt cgttaaaatg cgcacaatac tggccacaaa aagaagaaaa agagatgatc
     1681 tttgaagaca caaatttgaa attaacattg atctctgaag atatcaagtc atattataca
     1741 gtgcgacagc tagaattgga aaaccttaca acccaagaaa ctcgagagat cttacatttc
     1801 cactatacca catggcctga ctttggagtc cctgaatcac cagcctcatt cttgaacttt
     1861 cttttcaaag tccgagagtc agggtcactc agcccggagc acgggcccgt tgtggtgcac
     1921 tgcagtgcag gcatcggcag gtctggaacc ttctgtctgg ctgatacctg cctcttgctg
     1981 atggacaaga ggaaagaccc ttcttccgtt gatatcaaga aagtgctgtt agaaatgagg
     2041 aagtttcgga tggggctgat ccagacagcc gaccagctgc gcttctccta cctggctgtg
     2101 atcgaaggtg ccaaattcat catgggggac tcttccgtgc aggatcagtg gaaggagctt
     2161 tcccacgagg acctggagcc cccacccgag catatccccc cacctccccg gccacccaaa
     2221 cgaatcctgg agccacacaa tgggaaatgc agggagttct tcccaaatta gggtaccctc
     2281 gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact
     2341 agcgcagctt aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt
     2401 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     2461 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2521 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2761 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2821 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2881 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttctggc ggcacgatgg
     2941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3901 actcttcctt tttcaatcat gattgaagca tttatcaggg ttattgtctc atgagcggat
     3961 acatatttga atgtatttag aaaaataaac aaataggtca tgaccaaaat cccttaacgt
     4021 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4081 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4141 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4201 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4261 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4321 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4381 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4441 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4501 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4561 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4621 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4681 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4741 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4801 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4861 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     4921 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4981 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5041 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5101 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     5161 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5221 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5281 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     5341 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     5401 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     5461 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     5521 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     5581 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     5641 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     5701 gcctagccgg gtcctcaacg acaggagcac gatcatgcta gtcatgcccc gcgcccaccg
     5761 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     5821 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5881 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5941 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     6001 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     6061 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     6121 ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     6181 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     6241 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     6301 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     6361 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     6421 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     6481 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     6541 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     6601 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     6661 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     6721 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     6781 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     6841 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     6901 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     6961 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     7021 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     7081 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     7141 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     7201 gcccgatctt ccccatcggt gatgtc
//
