LOCUS       Exported                7848 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of myc-his tagged AXL kinase dead (K558R) 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    AXL KD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7848)
  AUTHORS   Avilla E, Guarino V, Visciano C, Liotti F, Svelto M, Krishnamoorthy 
            G, Franco R, Melillo RM
  TITLE     Activation of TYRO3/AXL tyrosine kinase receptors in thyroid cancer.
  JOURNAL   Cancer Res. 2011 Mar 1;71(5):1792-804. doi: 
            10.1158/0008-5472.CAN-10-2186. Epub 2011 Feb 22.
  PUBMED    21343401
REFERENCE   2  (bases 1 to 7848)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7848
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(40..59)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        231..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        611..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    816..834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    837..855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     865..889
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3767..3796
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3812..3829
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3852..3869)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3858..4082
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4128..4556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4215..4234)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4425..4446
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4565..4585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4570..4899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4750..4885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4812..4831
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4947..4994
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5013..5387
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5517..5638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5554..5573)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5608..5627
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5687..5703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5687..5703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5700..5722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5711..5727
                     /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(5735..5765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5780..5801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5918..5935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6089..6677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6169..6188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6848..7708)
                     /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     7471..7490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7709..7813)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
       61 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
      121 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      181 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata gagatctccc
      841 tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg
      901 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      961 agcgtttaaa cttaagcttg gtaccgagct cggatccagc ccaacaactt ctgaggaaag
     1021 tttggcaccc atggcgtggc ggtgccccag gatgggcagg gtcccgctgg cctggtgctt
     1081 ggcgctgtgc ggctgggcgt gcatggcccc caggggcacg caggctgaag aaagtccctt
     1141 cgtgggcaac ccagggaata tcacaggtgc ccggggactc acgggcaccc ttcggtgtca
     1201 gctccaggtt cagggagagc cccccgaggt acattggctt cgggatggac agatcctgga
     1261 gctcgcggac agcacccaga cccaggtgcc cctgggtgag gatgaacagg atgactggat
     1321 agtggtcagc cagctcagaa tcacctccct gcagctttcc gacacgggac agtaccagtg
     1381 tttggtgttt ctgggacatc agaccttcgt gtcccagcct ggctatgttg ggctggaggg
     1441 cttgccttac ttcctggagg agcccgaaga caggactgtg gccgccaaca cccccttcaa
     1501 cctgagctgc caagctcagg gacccccaga gcccgtggac ctactctggc tccaggatgc
     1561 tgtccccctg gccacggctc caggtcacgg cccccagcgc agcctgcatg ttccagggct
     1621 gaacaagaca tcctctttct cctgcgaagc ccataacgcc aagggggtca ccacatcccg
     1681 cacagccacc atcacagtgc tcccccagca gccccgtaac ctccacctgg tctcccgcca
     1741 acccacggag ctggaggtgg cttggactcc aggcctgagc ggcatctacc ccctgaccca
     1801 ctgcaccctg caggctgtgc tgtcagacga tgggatgggc atccaggcgg gagaaccaga
     1861 ccccccagag gagcccctca cctcgcaagc atccgtgccc ccccatcagc ttcggctagg
     1921 cagcctccat cctcacaccc cttatcacat ccgcgtggca tgcaccagca gccagggccc
     1981 ctcatcctgg acccactggc ttcctgtgga gacgccggag ggagtgcccc tgggcccccc
     2041 tgagaacatt agtgctacgc ggaatgggag ccaggccttc gtgcattggc aagagccccg
     2101 ggcgcccctg cagggtaccc tgttagggta ccggctggcg tatcaaggcc aggacacccc
     2161 agaggtgcta atggacatag ggctaaggca agaggtgacc ctggagctgc agggggacgg
     2221 gtctgtgtcc aatctgacag tgtgtgtggc agcctacact gctgctgggg atggaccctg
     2281 gagcctccca gtacccctgg aggcctggcg cccagggcaa gcacagccag tccaccagct
     2341 ggtgaaggaa ccttcaactc ctgccttctc gtggccctgg tggtatgtac tgctaggagc
     2401 agtcgtggcc gctgcctgtg tcctcatctt ggctctcttc cttgtccacc ggcgaaagaa
     2461 ggagacccgt tatggagaag tgtttgaacc aacagtggaa agaggtgaac tggtagtcag
     2521 gtaccgcgtg cgcaagtcct acagtcgtcg gaccactgaa gctaccttga acagcttggg
     2581 catcagtgaa gagctgaagg agaagctgcg ggatgtgatg gtggaccggc acaaggtggc
     2641 cctggggaag actctgggag agggagagtt tggagctgtg atggaaggcc agctcaacca
     2701 ggacgactcc atcctcaagg tggctgtgag gacgatgaag attgccatct gcacgaggtc
     2761 agagctggag gatttcctga gtgaagcggt ctgcatgaag gaatttgacc atcccaacgt
     2821 catgaggctc atcggtgtct gtttccaggg ttctgaacga gagagcttcc cagcacctgt
     2881 ggtcatctta cctttcatga aacatggaga cctacacagc ttcctcctct attcccggct
     2941 cggggaccag ccagtgtacc tgcccactca gatgctagtg aagttcatgg cagacatcgc
     3001 cagtggcatg gagtatctga gtaccaagag attcatacac cgggacctgg cggccaggaa
     3061 ctgcatgctg aatgagaaca tgtccgtgtg tgtggcggac ttcgggctct ccaagaagat
     3121 ctacaatggg gactactacc gccagggacg tatcgccaag atgccagtca agtggattgc
     3181 cattgagagt ctagctgacc gtgtctacac cagcaagagc gatgtgtggt ccttcggggt
     3241 gacaatgtgg gagattgcca caagaggcca aaccccatat ccgggcgtgg agaacagcga
     3301 gatttatgac tatctgcgcc ggggaaatcg cctgaagcag cctgcggact gtctggatgg
     3361 actgtatgcc ttgatgtcgc ggtgctggga gctaaatccc caggaccggc caagttttac
     3421 agagctgcgg gaagatttgg agaacacact gaaggccttg cctcctgccc aggagcctga
     3481 cgaaatcctc tatgtcaaca tggatgaggg tggaggttat cctgaacccc ctggagctgc
     3541 aggaggagct gaccccccaa cccagccaga ccctaaggat tcctgtagct gcctcactgc
     3601 ggctgaggtc catcctgctg gacgctatgt cctctgccct tccacaaccc ctagccccgc
     3661 tcagcctgct gataggggct ccccagcagc cccagggcag gaggatggtg ccgaattctg
     3721 cagatatcca gcacagtggc ggccgctcga gtctagaggg cccttcgaac aaaaactcat
     3781 ctcagaagag gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttaaac
     3841 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3901 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3961 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4021 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4081 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4141 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4201 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4261 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4321 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4381 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4441 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4501 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4561 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4621 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4681 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4741 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4801 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4861 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4921 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     4981 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     5041 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     5101 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     5161 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     5221 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     5281 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     5341 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     5401 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5461 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5521 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5581 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5641 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5761 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5821 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5881 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7801 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacg
//
