LOCUS       sequence-115157-        7811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AXL on cistronic transcript with puro resistance in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7811)
  AUTHORS   Meyer AS, Zweemer AJ, Lauffenburger DA
  TITLE     The AXL Receptor is a Sensor of Ligand Spatial Heterogeneity.
  JOURNAL   Cell Syst. 2015 Jul 29;1(1):25-36.
  PUBMED    26236777
REFERENCE   2  (bases 1 to 7811)
  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"; journalName: "Cell 
            Syst."; date: "2015-07-29"; volume: "1(1)"; pages: "25-36"
COMMENT     Sequence Label: pIRESpuro2 AXL
FEATURES             Location/Qualifiers
     source          1..7811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      933..942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3670..3899
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3882..3901
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3957..4531,4532..4534,4535..4543)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4124..4141)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4351..4370
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4563..5162
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(4563..4582)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5059..5079
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5298..5315)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5304..5528
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5575..5594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5654..5670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5654..5670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5667..5689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5678..5694
                     /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(5702..5708,5709..5726,5727..5732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5747..5768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5885..5902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6056..6644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6136..6155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6815..7606,7607..7675))
                     /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     7438..7457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7676..7780)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgccaccat ggcgtggcgg tgccccagga
      961 tgggcagggt cccgctggcc tggtgcttgg cgctgtgcgg ctgggcgtgc atggccccca
     1021 ggggcacgca ggctgaagaa agtcccttcg tgggcaaccc agggaatatc acaggtgccc
     1081 ggggactcac gggcaccctt cggtgtcagc tccaggttca gggagagccc cccgaggtac
     1141 attggcttcg ggatggacag atcctggagc tcgcggacag cacccagacc caggtgcccc
     1201 tgggtgagga tgaacaggat gactggatag tggtcagcca gctcagaatc acctccctgc
     1261 agctttccga cacgggacag taccagtgtt tggtgtttct gggacatcag accttcgtgt
     1321 cccagcctgg ctatgttggg ctggagggct tgccttactt cctggaggag cccgaagaca
     1381 ggactgtggc cgccaacacc cccttcaacc tgagctgcca agctcaggga cccccagagc
     1441 ccgtggacct actctggctc caggatgctg tccccctggc cacggctcca ggtcacggcc
     1501 cccagcgcag cctgcatgtt ccagggctga acaagacatc ctctttctcc tgcgaagccc
     1561 ataacgccaa gggggtcacc acatcccgca cagccaccat cacagtgctc ccccagcagc
     1621 cccgtaacct ccacctggtc tcccgccaac ccacggagct ggaggtggct tggactccag
     1681 gcctgagcgg catctacccc ctgacccact gcaccctgca ggctgtgctg tcagacgatg
     1741 ggatgggcat ccaggcggga gaaccagacc ccccagagga gcccctcacc tcgcaagcat
     1801 ccgtgccccc ccatcagctt cggctaggca gcctccatcc tcacacccct tatcacatcc
     1861 gcgtggcatg caccagcagc cagggcccct catcctggac ccactggctt cctgtggaga
     1921 cgccggaggg agtgcccctg ggcccccctg agaacattag tgctacgcgg aatgggagcc
     1981 aggccttcgt gcattggcaa gagccccggg cgcccctgca gggtaccctg ttagggtacc
     2041 ggctggcgta tcaaggccag gacaccccag aggtgctaat ggacataggg ctaaggcaag
     2101 aggtgaccct ggagctgcag ggggacgggt ctgtgtccaa tctgacagtg tgtgtggcag
     2161 cctacactgc tgctggggat ggaccctgga gcctcccagt acccctggag gcctggcgcc
     2221 cagtgaagga accttcaact cctgccttct cgtggccctg gtggtatgta ctgctaggag
     2281 cagtcgtggc cgctgcctgt gtcctcatct tggctctctt ccttgtccac cggcgaaaga
     2341 aggagacccg ttatggagaa gtgtttgaac caacagtgga aagaggtgaa ctggtagtca
     2401 ggtaccgcgt gcgcaagtcc tacagtcgtc ggaccactga agctaccttg aacagcctgg
     2461 gcatcagtga agagctgaag gagaagctgc gggatgtgat ggtggaccgg cacaaggtgg
     2521 ccctggggaa gactctggga gagggagagt ttggagctgt gatggaaggc cagctcaacc
     2581 aggacgactc catcctcaag gtggctgtga agacgatgaa gattgccatc tgcacgaggt
     2641 cagagctgga ggatttcctg agtgaagcgg tctgcatgaa ggaatttgac catcccaacg
     2701 tcatgaggct catcggtgtc tgtttccagg gttctgaacg agagagcttc ccagcacctg
     2761 tggtcatctt acctttcatg aaacatggag acctacacag cttcctcctc tattcccggc
     2821 tcggggacca gccagtgtac ctgcccactc agatgctagt gaagttcatg gcagacatcg
     2881 ccagtggcat ggagtatctg agtaccaaga gattcataca ccgggacctg gcggccagga
     2941 actgcatgct gaatgagaac atgtccgtgt gtgtggcgga cttcgggctc tccaagaaga
     3001 tctacaatgg ggactactac cgccagggac gtatcgccaa gatgccagtc aagtggattg
     3061 ccattgagag tctagctgac cgtgtctaca ccagcaagag cgatgtgtgg tccttcgggg
     3121 tgacaatgtg ggagattgcc acaagaggcc aaaccccata tccgggcgtg gagaacagcg
     3181 agatttatga ctatctgcgc cggggaaatc gcctgaagca gcctgcggac tgtctggatg
     3241 gactgtatgc cttgatgtcg cggtgctggg agctaaatcc ccaggaccgg ccaagtttta
     3301 cagagctgcg ggaagatttg gagaacacac tgaaggcctt gcctcctgcc caggagcctg
     3361 acgaaatcct ctatgtcaac atggatgagg gtggaggtta tcctgaaccc cctggagctg
     3421 caggaggagc tgacccccca acccagccag accctaagga ttcctgtagc tgcctcactg
     3481 cggctgaggt ccatcctgct ggacgctatg tcctctgccc ttccacaacc cctagccccg
     3541 ctcagcctgc tgataggggc tccccagcag ccccagggca ggaggatggt gccccggatc
     3601 cgcggccgca tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca
     3661 gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag
     3721 tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg
     3781 ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc
     3841 ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg
     3901 tgtccactcc caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc
     3961 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     4021 tttgtctata tgtgattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     4081 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     4141 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     4201 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     4261 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     4321 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     4381 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     4441 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     4501 gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccacaagg agacgacctt
     4561 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     4621 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     4681 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4741 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4801 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4861 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4921 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4981 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5041 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5101 ccgacgtcga gtgcccgaag gaccgcgcga cctggtgcat gacccgcaag cccggtgcct
     5161 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg
     5221 accgaagccg acccgggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgactc
     5281 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     5341 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5401 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5461 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     5521 ctctatggct tctgaggcgg aaagaaccag ctggggctcg agtgcattct agttgtggtt
     5581 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5641 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5701 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5761 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5821 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6241 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6481 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7801 cacctgacgt c
//
