LOCUS       sequence-388271-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MerTK Kinase Domain in E. coli and surface entropy 
            reduction mutant for crystallization, coexpress PTPN1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  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 7232)
  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_KRmutant-PTPN1
FEATURES             Location/Qualifiers
     source          1..7232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        1101..1119
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1120..1144
                     /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             2198..2242
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2280..2298)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2294..2341
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2378..2833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2465..2484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2675..2696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2948..3739,3740..3808))
                     /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     3571..3590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3809..3901)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3982..4570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4471..4490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4724..4741
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4756..4895
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4821..4840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4981..5003
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4997..5188)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5760..6842)
                     /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     6804..6823
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6843..6920)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7126..7145
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg ttccgcgtgg tagtcatatg gaggaactac aaaataaact agaagatgtt
      181 gtgattgaca ggaatcttct aattcttgga agaattctgg gtgaaggaga gtttgggtct
      241 gtaatggaag gaaatcttaa gcaggaagat gggacctctc tgaaagtggc agtgaagacc
      301 atgaagttgg acaactcttc acagcgggag atcgaggagt ttctcagtga ggcagcgtgc
      361 atgaaagact tcagccaccc aaatgtcatt cgacttctag gtgtgtgtat agaaatgagc
      421 tctcaaggca tcccaaagcc catggtaatt ttacccttca tgaaatacgg ggacctgcat
      481 acttacttac tttattcccg attggagaca ggaccaaggc atattcctct gcagacacta
      541 ttgaggttca tggtggatat tgccctggga atggagtatc tgagcaacag gaattttctt
      601 catcgagatt tagctgctcg aaactgcatg ttgcgagatg acatgactgt ctgtgttgcg
      661 gacttcggcc tctctaagaa gatttacagt ggcgattatt accgccaagg ccgcattgct
      721 aagatgcctg ttaaatggat cgccatagaa agtcttgcag accgagtcta cacaagtaaa
      781 agtgatgtgt gggcatttgg cgtgaccatg tgggaaatag ctacgcgggg aatgactccc
      841 tatcctgggg tccagaacca tgagatgtat gactatcttc tccatggcca caggttgaag
      901 cagcccgaag actgcctgga tgaactgtat gaaataatgt actcttgctg gagaaccgat
      961 cccttagacc gccccacctt ttcagtattg aggctgcagc tagaaagact cttagaaagt
     1021 ttgcctgacg tttgagcggc cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta
     1081 cacggccgca taatcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     1141 ttccccatct tagtatatta gttaagtata agaaggagat atacatatgg agatggaaaa
     1201 ggagttcgag cagatcgaca agtccgggag ctgggcggcc atttaccagg atatccgaca
     1261 tgaagccagt gacttcccat gtagagtggc caagcttcct aagaacaaaa accgaaatag
     1321 gtacagagac gtcagtccct ttgaccatag tcggattaaa ctacatcaag aagataatga
     1381 ctatatcaac gctagtttga taaaaatgga agaagcccaa aggagttaca ttcttaccca
     1441 gggccctttg cctaacacat gcggtcactt ttgggagatg gtgtgggagc agaaaagcag
     1501 gggtgtcgtc atgctcaaca gagtgatgga gaaaggttcg ttaaaatgcg cacaatactg
     1561 gccacaaaaa gaagaaaaag agatgatctt tgaagacaca aatttgaaat taacattgat
     1621 ctctgaagat atcaagtcat attatacagt gcgacagcta gaattggaaa accttacaac
     1681 ccaagaaact cgagagatct tacatttcca ctataccaca tggcctgact ttggagtccc
     1741 tgaatcacca gcctcattct tgaactttct tttcaaagtc cgagagtcag ggtcactcag
     1801 cccggagcac gggcccgttg tggtgcactg cagtgcaggc atcggcaggt ctggaacctt
     1861 ctgtctggct gatacctgcc tcttgctgat ggacaagagg aaagaccctt cttccgttga
     1921 tatcaagaaa gtgctgttag aaatgaggaa gtttcggatg gggctgatcc agacagccga
     1981 ccagctgcgc ttctcctacc tggctgtgat cgaaggtgcc aaattcatca tgggggactc
     2041 ttccgtgcag gatcagtgga aggagctttc ccacgaggac ctggagcccc cacccgagca
     2101 tatcccccca cctccccggc cacccaaacg aatcctggag ccacacaatg ggaaatgcag
     2161 ggagttcttc ccaaattagg gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt
     2221 tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac
     2281 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2341 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     2401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2521 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2581 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2641 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2701 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2761 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2821 acgtttacaa tttctggcgg cacgatggca tgagattatc aaaaaggatc ttcacctaga
     2881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatcatga ttgaagcatt
     3841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3901 ataggtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3961 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4021 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4081 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4141 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4201 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4261 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4321 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4381 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4441 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4501 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4561 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4621 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4681 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4741 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4801 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4861 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4921 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4981 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     5041 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     5101 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     5161 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     5221 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     5281 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     5341 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     5401 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     5461 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     5521 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     5581 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     5641 tcatgctagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     5701 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     5761 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5821 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5881 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5941 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     6001 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     6061 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     6121 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     6181 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     6241 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     6301 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     6361 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     6421 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     6481 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     6541 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     6601 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     6661 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     6721 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     6781 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     6841 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6901 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6961 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     7021 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     7081 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     7141 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     7201 gaggatcgag atcgatctcg atcccgcgaa at
//
