LOCUS Exported 7811 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses AXL on cistronic transcript with puro resistance in mammalian cells. ACCESSION . VERSION . KEYWORDS pIRESpuro2 AXL 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7811 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 3957..4530 /label=IRES /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(5702..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(6815..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 //