LOCUS       sequence-454328-        8021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimeric contruct having N-terminal human Tyro3 and 
            C-terminal human Interferon Gamma Receptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  AUTHORS   Tsou WI, Nguyen KQ, Calarese DA, Garforth SJ, Antes AL, Smirnov SV, 
            Almo SC, Birge RB, Kotenko SV
  TITLE     Receptor tyrosine kinases, TYRO3, AXL, and MER, demonstrate distinct
            patterns and complex regulation of ligand-induced activation.
  JOURNAL   J Biol Chem. 2014 Sep 12;289(37):25750-63. doi: 
            10.1074/jbc.M114.569020. Epub 2014 Jul 29.
  PUBMED    25074926
REFERENCE   2  (bases 1 to 8021)
  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: 
            "10.1074/jbc.M114.569020"; journalName: "J Biol Chem"; date: 
            "2014-09-12- 12"; volume: "289"; issue: "37"; pages: "25750-63"
COMMENT     Sequence Label: pEF2-FL-hTyro_gR1
FEATURES             Location/Qualifiers
     source          1..8021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(139..158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      952..1540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1441..1460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1694..1711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1828..1849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1864..1869,1870..1887,1888..1894)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1902..1918
                     /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)."
     primer_bind     1907..1929
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1926..1942
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1926..1942
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1991..2112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2002..2021)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2056..2075
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2286..3080)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2397..2416)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3007..3026
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3147..3476)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3161..3296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3215..3234)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3461..3481
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3490..3918)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3600..3621)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3812..3831
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3964..4188
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4177..4194
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        4214..4232
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4214..4231
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(6156..6179)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(6277..7455)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6286..7224
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6309..6329)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     7743..7762
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7837..7941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7942..8010,8011..8021,1..781)
                     /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"
ORIGIN
        1 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
       61 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      121 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
      181 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      241 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      301 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
      361 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
      421 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
      481 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
      541 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      601 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
      661 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
      721 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
      781 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
      841 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
      901 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
      961 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1021 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1081 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1141 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1201 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1261 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1321 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1381 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1441 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1501 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1561 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1621 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1681 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     1741 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     1801 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     1861 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     1921 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat
     1981 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2041 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2101 aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     2161 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     2221 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     2281 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     2341 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     2401 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     2461 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     2521 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     2581 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     2641 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     2701 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     2761 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     2821 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     2881 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     2941 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     3001 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     3061 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatccgaaa
     3121 atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa aaagcctcct
     3181 cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata aataaaaaaa
     3241 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     3301 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     3361 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     3421 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     3481 cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3541 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3601 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3661 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3721 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3781 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3841 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3901 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag
     3961 aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct
     4021 tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg
     4081 caatttcctc attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag
     4141 gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc
     4201 agcgagctct agcatttagg tgacactata gaatagggcc ctctagtagg aattcatgaa
     4261 aattctttgg aatcttctgt tggtctataa ccaatcaagg actctttacc gctatcatcc
     4321 acaagtagat ccactagcac atgtggttta tcataaccaa aggaggtggg ggcttttatt
     4381 acggttatga gctcttgtcc ttctgttttt atttcacctt tattatttgg gggaaattct
     4441 gagtcagata aggagctatg tgattccaga cagctggaat cagtatcaaa accatttctg
     4501 gagtgatcac tctcagaaca atttctggag tgatacgagt tcaaagcgat gctgccaggt
     4561 tcagactggt tactacttaa aggtgaagaa ctctctctct ctattggagt cagatggctg
     4621 cccgggacca cgtcaggaat attttcttca gtagtcacca cttctgttat actagaaagt
     4681 tcttctgtat gttccacttt tcctggattg tcttcggtat gcatgcctgg aactgttgct
     4741 ggagacaacg gctcttcaca gaccacctcc ttttctaagg aaaatggctg gtatgacgtg
     4801 atgagtgata catattttga ttcaggtttt gtctctaaag tagcacttct taccacagag
     4861 atcaaggact tgggtaatat tatgcttttt tccttcaatg gattaatttt cttaatataa
     4921 aaacagatga ataccaggct aagcactaga aagagtagta aagcagcaac aactggcgct
     4981 agccaggatg tgcggctgtg aggagggccc tgctggcctg cacggtcatg agaagagacc
     5041 accagtggct gactccaggg tccacagcca actgcattgg agacgcacac acgtacgatc
     5101 aggtcctttt ggggatccca gcctgtcaaa ttggccctgg tcccctccac tgtcagctca
     5161 tcctgggttc cattgtcttg aacccaggac agtttgtagg gtcccagggg gccttccaaa
     5221 ggggcctcgg ggatcacttc ttcccactcc aagatgaggc ctgaatctgt gcggatggca
     5281 tggaggtttt ggggagcgct ggctggggct agacccttgg tctgaaaggg cacccagtca
     5341 gcatagggag agggccccaa ggcattggca cagcgcaccc tgaggctgta gttggtggca
     5401 ggcaccaggt cccggagcag gcaggtaaag gggggcacag ggaccacaac agccaggact
     5461 tcccagcctc ctggggcctg tgtcacctga actgtacagg actgtagcag agctcggcca
     5521 tcagcacctg gcatccaggc cacactagcg ttgctgctgg aaagctttgt cacggtgatg
     5581 ttgaaggggg ctgcaggcag tgcttgaagg tgaacagtgg ctgtgcgaga agaggccagg
     5641 ccttttaggt tgtgagcttc acaggaaaac atggtgctct gggtcacccc tgttacattt
     5701 aaaacagatg gagagggagc gggtcccccg atcttcgtag ttcctctcca ccagacaatg
     5761 gtaacaggtt cagggggacc cacagcctca caagacagtt ggaaaggggc attgggtggc
     5821 actgccagat cttttggctc cactgtgaaa aatggcacac cttctaccgt gagccacact
     5881 ggctgggaga tctcggtttc acccccatcc tccacctggc accagtaccg gccggcgtca
     5941 gagcgctcca ctgacttcag gctgaggaag ccgatccagt gctgctcgct gactgggatg
     6001 tacaactggt ccaagttctg gaccacagcc ccatccttca cccactggat gtcaggctcc
     6061 tccatcccct ccacactgca gttgagcttc accggctgcc cctgagacac tgtcagcttc
     6121 accggggctc ccatgagctt cagaccgtga tccgccttgt catcgtcgtc cttgtagtcg
     6181 gcggcggcgg cggcgaagac tccgagcagc agcagcagcg accacagcag cgtcggtcgc
     6241 atggccccga tccgagctcg gtaccaagct aattcctcac gacacctgaa atggaagaaa
     6301 aaaactttga accactgtct gaggcttgag aatgaaccaa gatccaaact caaaaagggc
     6361 aaattccaag gagaattaca tcaagtgcca agctggccta acttcagtct ccacccactc
     6421 agtgtgggga aactccatcg cataaaaccc ctccccccaa cctaaagacg acgtactcca
     6481 aaagctcgag aactaatcga ggtgcctgga cggcgcccgg tactccgtgg agtcacatga
     6541 agcgacggct gaggacggaa aggccctttt cctttgtgtg ggtgactcac ccgcccgctc
     6601 tcccgagcgc cgcgtcctcc attttgagct ccctgcagca gggccgggaa gcggccatct
     6661 ttccgctcac gcaactggtg ccgaccgggc cagccttgcc gcccagggcg gggcgataca
     6721 cggcggcgcg aggccaggca ccagagcagg ccggccagct tgagactacc cccgtccgat
     6781 tctcggtggc cgcgctcgca ggccccgcct cgccgaacat gtgcgctggg acgcacgggc
     6841 cccgtcgccg cccgcggccc caaaaaccga aataccagtg tgcagatctt ggcccgcatt
     6901 tacaagacta tcttgccaga aaaaaagcgt cgcagcaggt catcaaaaat tttaaatggc
     6961 tagagactta tcgaaagcag cgagacaggc gcgaaggtgc caccagattc gcacgcggcg
     7021 gccccagcgc ccaggccagg cctcaactca agcacgaggc gaaggggctc cttaagcgca
     7081 aggcctcgaa ctctcccacc cacttccaac ccgaagctcg ggatcaagaa tcacgtactg
     7141 cagccaggtg gaagtaattc aaggcacgca agggccataa cccgtaaaga ggccaggccc
     7201 gcgggaacca cacacggcac ttacctgtgt tctggcggca aacccgttgc gaaaaagaac
     7261 gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag
     7321 ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg caccggttca
     7381 attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccactg
     7441 acgggcaccg gagccaattc ccactccttt caagacctag aaggtccatt agctgcaaag
     7501 attcctctct gtttaaaact ttatccatct ttgcaaagct ttttgcaaaa gcctagtcaa
     7561 taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac
     7621 cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt
     7681 gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta
     7741 tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat
     7801 ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     7861 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7921 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7981 tttgcggcat tttgccttcc tgtttttgct cacccagaaa c
//
