LOCUS       Exported                7333 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of Flag-tagged AXL.
ACCESSION   .
VERSION     .
KEYWORDS    pFLAG AXL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7333)
  AUTHORS   Krishnamoorthy GP, Guida T, Alfano L, Avilla E, Santoro M, 
            Carlomagno F, Melillo RM
  TITLE     Molecular mechanism of 17-allylamino-17-demethoxygeldanamycin 
            (17-AAG)-induced AXL receptor tyrosine kinase degradation.
  JOURNAL   J Biol Chem. 2013 Jun 14;288(24):17481-94. doi: 
            10.1074/jbc.M112.439422. Epub 2013 Apr 29.
  PUBMED    23629654
REFERENCE   2  (bases 1 to 7333)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7333
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     114..136
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     128..145
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     129..145
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        306..685
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        686..889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     840..860
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3631..3654
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3662..4284
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3861..3880)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4308..4328)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4313..4642
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4493..4628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4555..4574
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4680..4699)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4681..4699)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4713..4729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4713..4729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4726..4748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4737..4753
                     /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(4761..4791)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4806..4827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4944..4961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5115..5703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5195..5214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5874..6734)
                     /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     6497..6516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6735..6839)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6866..7321
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6953..6972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7163..7184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
       61 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      121 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
      181 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
      241 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
      301 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      361 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      421 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      481 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      541 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
      601 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
      661 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
      721 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      781 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
      841 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      901 ccgtcagaat tcacttctga ggaaagtttg gcacccatgg cgtggcggtg ccccaggatg
      961 ggcagggtcc cgctggcctg gtgcttggcg ctgtgcggct gggcgtgcat ggcccccagg
     1021 ggcacgcagg ctgaagaaag tcccttcgtg ggcaacccag ggaatatcac aggtgcccgg
     1081 ggactcacgg gcacccttcg gtgtcagctc caggttcagg gagagccccc cgaggtacat
     1141 tggcttcggg atggacagat cctggagctc gcggacagca cccagaccca ggtgcccctg
     1201 ggtgaggatg aacaggatga ctggatagtg gtcagccagc tcagaatcac ctccctgcag
     1261 ctttccgaca cgggacagta ccagtgtttg gtgtttctgg gacatcagac cttcgtgtcc
     1321 cagcctggct atgttgggct ggagggcttg ccttacttcc tggaggagcc cgaagacagg
     1381 actgtggccg ccaacacccc cttcaacctg agctgccaag ctcagggacc cccagagccc
     1441 gtggacctac tctggctcca ggatgctgtc cccctggcca cggctccagg tcacggcccc
     1501 cagcgcagcc tgcatgttcc agggctgaac aagacatcct ctttctcctg cgaagcccat
     1561 aacgccaagg gggtcaccac atcccgcaca gccaccatca cagtgctccc ccagcagccc
     1621 cgtaacctcc acctggtctc ccgccaaccc acggagctgg aggtggcttg gactccaggc
     1681 ctgagcggca tctaccccct gacccactgc accctgcagg ctgtgctgtc agacgatggg
     1741 atgggcatcc aggcgggaga accagacccc ccagaggagc ccctcacctc gcaagcatcc
     1801 gtgccccccc atcagcttcg gctaggcagc ctccatcctc acacccctta tcacatccgc
     1861 gtggcatgca ccagcagcca gggcccctca tcctggaccc actggcttcc tgtggagacg
     1921 ccggagggag tgcccctggg cccccctgag aacattagtg ctacgcggaa tgggagccag
     1981 gccttcgtgc attggcaaga gccccgggcg cccctgcagg gtaccctgtt agggtaccgg
     2041 ctggcgtatc aaggccagga caccccagag gtgctaatgg acatagggct aaggcaagag
     2101 gtgaccctgg agctgcaggg ggacgggtct gtgtccaatc tgacagtgtg tgtggcagcc
     2161 tacactgctg ctggggatgg accctggagc ctcccagtac ccctggaggc ctggcgccca
     2221 gggcaagcac agccagtcca ccagctggtg aaggaacctt caactcctgc cttctcgtgg
     2281 ccctggtggt atgtactgct aggagcagtc gtggccgctg cctgtgtcct catcttggct
     2341 ctcttccttg tccaccggcg aaagaaggag acccgttatg gagaagtgtt tgaaccaaca
     2401 gtggaaagag gtgaactggt agtcaggtac cgcgtgcgca agtcctacag tcgtcggacc
     2461 actgaagcta ccttgaacag cttgggcatc agtgaagagc tgaaggagaa gctgcgggat
     2521 gtgatggtgg accggcacaa ggtggccctg gggaagactc tgggagaggg agagtttgga
     2581 gctgtgatgg aaggccagct caaccaggac gactccatcc tcaaggtggc tgtgaagacg
     2641 atgaagattg ccatctgcac gaggtcagag ctggaggatt tcctgagtga agcggtctgc
     2701 atgaaggaat ttgaccatcc caacgtcatg aggctcatcg gtgtctgttt ccagggttct
     2761 gaacgagaga gcttcccagc acctgtggtc atcttacctt tcatgaaaca tggagaccta
     2821 cacagcttcc tcctctattc ccggctcggg gaccagccag tgtacctgcc cactcagatg
     2881 ctagtgaagt tcatggcaga catcgccagt ggcatggagt atctgagtac caagagattc
     2941 atacaccggg acctggcggc caggaactgc atgctgaatg agaacatgtc cgtgtgtgtg
     3001 gcggacttcg ggctctccaa gaagatctac aatggggact actaccgcca gggacgtatc
     3061 gccaagatgc cagtcaagtg gattgccatt gagagtctag ctgaccgtgt ctacaccagc
     3121 aagagcgatg tgtggtcctt cggggtgaca atgtgggaga ttgccacaag aggccaaacc
     3181 ccatatccgg gcgtggagaa cagcgagatt tatgactatc tgcgccgggg aaatcgcctg
     3241 aagcagcctg cggactgtct ggatggactg tatgccttga tgtcgcggtg ctgggagcta
     3301 aatccccagg accggccaag ttttacagag ctgcgggaag atttggagaa cacactgaag
     3361 gccttgcctc ctgcccagga gcctgacgaa atcctctatg tcaacatgga tgagggtgga
     3421 ggttatcctg aaccccctgg agctgcagga ggagctgacc ccccaaccca gccagaccct
     3481 aaggattcct gtagctgcct cactgcggct gaggtccatc ctgctggacg ctatgtcctc
     3541 tgcccttcca caacccctag ccccgctcag cctgctgata ggggctcccc agcagcccca
     3601 gggcaggagg atggtgccgg atccggtacc gattacaagg acgacgatga caagtagtac
     3661 tgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3721 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3781 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3841 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3901 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3961 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     4021 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     4081 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     4141 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca
     4201 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc
     4261 cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca
     4321 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4381 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4441 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4501 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4561 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4621 tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc
     4681 cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     4741 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4801 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4861 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4921 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5521 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6181 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6481 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6841 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6901 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6961 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     7021 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     7081 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     7141 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     7201 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     7261 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     7321 tccattcgcc att
//
