LOCUS       sequence-454318-        8111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a chimeric contruct having N-terminal human Axl and 
            C-terminal human Interferon Gamma Receptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8111)
  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 8111)
  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-hAxl_gR1
FEATURES             Location/Qualifiers
     source          1..8111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        268..1446
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          499..1437
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1394..1414
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1544..1567
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3581..3599)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3582..3599)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3619..3636)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3625..3849
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3895..4323
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3982..4001)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4192..4213
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4332..4352)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4337..4666
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4517..4652
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4579..4598
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4733..5527
                     /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(4787..4806)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5397..5416
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5701..5822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5738..5757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5792..5811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5871..5887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5871..5887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5884..5906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5895..5911
                     /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)."
     promoter        complement(join(5919..5925,5926..5943,5944..5949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5964..5985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6102..6119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6273..6861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6353..6372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7032..7823,7824..7892))
                     /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     7655..7674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7893..7997)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8072..8091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
       61 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      121 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag gcttttgcaa
      181 aaagctttgc aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc
      241 ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
      301 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
      361 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
      421 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
      481 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
      541 ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat
      601 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
      661 tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg
      721 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
      781 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
      841 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
      901 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
      961 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
     1021 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
     1081 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
     1141 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
     1201 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
     1261 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
     1321 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
     1381 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
     1441 tcgtgaggaa ttagcttggt accgagctcg gatcggggcc atgcgaccga cgctgctgtg
     1501 gtcgctgctg ctgctgctcg gagtcttcgc cgccgccgcc gccgactaca aggacgacga
     1561 tgacaaggcg gatcccgaag aaagtccctt cgtgggcaac ccagggaata tcacaggtgc
     1621 ccggggactc acgggcaccc ttcggtgtca gctccaggtt cagggagagc cccccgaggt
     1681 acattggctt cgggatggac agatcctgga gctcgcggac agcacccaga cccaggtgcc
     1741 cctgggtgag gatgaacagg atgactggat agtggtcagc cagctcagaa tcacctccct
     1801 gcagctttcc gacacgggac agtaccagtg tttggtgttt ctgggacatc agaccttcgt
     1861 gtcccagcct ggctatgttg ggctggaggg cttgccttac ttcctggagg agcccgaaga
     1921 caggactgtg gccgccaaca cccccttcaa cctgagctgc caagctcagg gacccccaga
     1981 gcccgtggac ctactctggc tccaggatgc tgtccccctg gccacggctc caggtcacgg
     2041 cccccagcgc agcctgcatg ttccagggct gaacaagaca tcctctttct cctgcgaagc
     2101 ccataacgcc aagggggtca ccacatcccg cacagccacc atcacagtgc tcccccagca
     2161 gccccgtaac ctccacctgg tctcccgcca acccacggag ctggaggtgg cttggactcc
     2221 aggcctgagc ggcatctacc ccctgaccca ctgcaccctg caggctgtgc tgtcagacga
     2281 tgggatgggc atccaggcgg gagaaccaga ccccccagag gagcccctca cctcgcaagc
     2341 atccgtgccc ccccatcagc ttcggctagg cagcctccat cctcacaccc cttatcacat
     2401 ccgcgtggca tgcaccagca gccagggccc ctcatcctgg acccactggc ttcctgtgga
     2461 gacgccggag ggagtgcccc tgggcccccc tgagaacatt agtgctacgc ggaatgggag
     2521 ccaggccttc gtgcattggc aagagccccg ggcgcccctg cagggtaccc tgttagggta
     2581 ccggctggcg tatcaaggcc aggacacccc agaggtgcta atggacatag ggctaaggca
     2641 agaggtgacc ctggagctgc agggggacgg gtctgtgtcc aatctgacag tgtgtgtggc
     2701 agcctacact gctgctgggg atggaccctg gagcctccca gtacccctgg aggcctggcg
     2761 cccagggcaa gcacagccag tccaccagct ggtgaaggaa ccttcaactc ctgccttctc
     2821 gtggccctgg ctagcgccag ttgttgctgc tttactactc tttctagtgc ttagcctggt
     2881 attcatctgt ttttatatta agaaaattaa tccattgaag gaaaaaagca taatattacc
     2941 caagtccttg atctctgtgg taagaagtgc tactttagag acaaaacctg aatcaaaata
     3001 tgtatcactc atcacgtcat accagccatt ttccttagaa aaggaggtgg tctgtgaaga
     3061 gccgttgtct ccagcaacag ttccaggcat gcataccgaa gacaatccag gaaaagtgga
     3121 acatacagaa gaactttcta gtataacaga agtggtgact actgaagaaa atattcctga
     3181 cgtggtcccg ggcagccatc tgactccaat agagagagag agttcttcac ctttaagtag
     3241 taaccagtct gaacctggca gcatcgcttt gaactcgtat cactccagaa attgttctga
     3301 gagtgatcac tccagaaatg gttttgatac tgattccagc tgtctggaat cacatagctc
     3361 cttatctgac tcagaatttc ccccaaataa taaaggtgaa ataaaaacag aaggacaaga
     3421 gctcataacc gtaataaaag cccccacctc ctttggttat gataaaccac atgtgctagt
     3481 ggatctactt gtggatgata gcggtaaaga gtccttgatt ggttatagac caacagaaga
     3541 ttccaaagaa ttttcatgaa ttcctactag agggccctat tctatagtgt cacctaaatg
     3601 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3661 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3721 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3781 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3841 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3901 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3961 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4021 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4081 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4141 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4201 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4261 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4321 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4381 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4501 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4561 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4621 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4681 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4741 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4801 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4861 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4921 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4981 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5041 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5101 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5161 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5221 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5281 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5341 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5401 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5461 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5521 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5581 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5641 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5821 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5881 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5941 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6001 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6061 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6241 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7141 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7981 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8041 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     8101 gccagtatct g
//
