LOCUS Exported 8101 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression of cell-surface protein extracellular domain fused to COMP5AP-AviTag-9xHis. Protein is secreted from cells.. ACCESSION . VERSION . KEYWORDS pD649-HAsp-IL1RL2-COMP5AP-AviTag-9xHis SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8101) AUTHORS Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC TITLE A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions. JOURNAL Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 10.1016/j.cell.2020.07.025. PUBMED 32822567 REFERENCE 2 (bases 1 to 8101) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8101 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 124..143 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 149..168 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 254..842 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 743..762 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 996..1013 /label=L4440 /note="L4440 vector, forward primer" primer_bind 1111..1128 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1112..1128 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1584..1600 /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)." enhancer 1767..2146 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2147..2350 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2301..2321 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2347..2371 /label=LNCX /note="Human CMV promoter, forward primer" promoter 2581..2599 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2581..2598 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 2612..2631 /label=pMT2-F /note="Synthetic intron, forward primer" CDS 3751..3774 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 5434..5478 /codon_start=1 /product="peptide tag that allows for enzymatic biotinylation" /label=AviTag(TM) /translation="GLNDIFEAQKIEWHE" CDS 5479..5505 /codon_start=1 /product="9xHis affinity tag" /label=9xHis /translation="HHHHHHHHH" primer_bind complement(5645..5662) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5651..5762 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(5880..5900) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5885..6214 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6065..6200 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6127..6146 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6262..6309 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6328..6927 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6328..6347) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6824..6844 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 7057..7178 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7094..7113) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7148..7167 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(7231..8091) /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 7854..7873 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8092..82) /gene="bla" /label=AmpR promoter ORIGIN 1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac 121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa 181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga 241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc 1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg 1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct 1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg 1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa 1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc 1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga 1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc 1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag 1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt 1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa 1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg 1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc 1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg 2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga 2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac 2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc 2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg 2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc 2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc 2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgccga tggctgcaag 2761 gacatcttta tgaagaacga gatcctgagc gccagccagc ctttcgcctt caactgcacc 2821 tttccaccta tcacctctgg cgaggtgtcc gtgacctggt acaagaacag cagcaagatc 2881 cccgtgtcca agatcatcca gagcagaatc caccaggacg agacatggat tctgtttctg 2941 cccatggaat ggggcgacag cggagtgtac cagtgcgtga tcaagggcag agacagctgc 3001 catagaatcc acgtgaacct gaccgtgttc gagaaacatt ggtgcgacac cagcatcggc 3061 ggactgccta atctgagcga cgagtacaag cagatcctgc acctgggcaa agacgacagc 3121 ctgacctgtc atctgcactt ccctaagagc tgcgtgctgg gccccatcaa gtggtacaag 3181 gactgcaacg agatcaaagg cgagcggttc accgtgctgg aaaccagact gctcgtgtcc 3241 aatgtgtccg ccgaggacag aggcaattac gcctgtcagg ccatcctgac acacagcggc 3301 aagcagtatg aggtgctgaa cggcatcacc gtgtccatca cagaaagagc cggatacggc 3361 ggcagcgtgc caaagatcat ctaccccaag aaccacagca tcgaggtgca gctgggcacc 3421 acactgatcg tggactgcaa tgtgaccgac accaaggaca acaccaacct gcggtgttgg 3481 agagtgaaca acaccctggt ggacgactac tacgacgaga gcaagagaat cagagaaggc 3541 gtggaaaccc acgtgtcctt cagagagcac aacctgtata ccgtgaatat caccttcctg 3601 gaagtgaaga tggaagatta cggcctgcct ttcatgtgcc atgccggcgt gtccacagcc 3661 tacatcattc tgcaactgcc cgctcctgac ttcagaggcg cgccggaaga tcaagttgat 3721 ccaagattga ttgatggcaa gggcggatct ctggaggtgc tgttccaggg accaggcgga 3781 agcggcaatc ccggcggcga tctggccccc cagatgctgc gcgagctgca ggagaccaat 3841 gccgccctgc aggacgtgcg cgagctgctg cgccaccgcg tgaaggagat caccttcctg 3901 aagaacaccg tgatggagtg cgacgcctgc ggtatgcagc cctctagagg ctccggctcc 3961 atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc cgaggccctg 4021 ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat catcttcctg 4081 ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg gcagaagaag 4141 gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt ggctctgtcc 4201 aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac ggcctacctg 4261 tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg ctttaaccag 4321 tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg 4381 aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc cggcacctac 4441 gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc ggcccgccag 4501 gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga cgtgatccta 4561 ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta cccagatgac 4621 tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg gctggcgaag 4681 cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc cctggacccg 4741 tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga gatccaccga 4801 gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg cctgctgagc 4861 aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca tggtcatcat 4921 gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc cattgagagg 4981 gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga ccactcccac 5041 gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct ggcccctggc 5101 aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc aggctatgtg 5161 ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc cgagtatcgg 5221 cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt ggcggtgttc 5281 gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt catagcgcac 5341 gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc gccccccgcc 5401 ggcaccaccg acgccgcgca cccgggtggc ggaggactga acgacatctt cgaggctcag 5461 aaaatcgaat ggcacgaaca tcaccatcac catcaccatc accattaagg ccggcctgta 5521 gtcactggca caacgcgtta cccggtaagc caatcgggta tacacggtcg tcatactgca 5581 gacagggttc ttctactttg caagatagtc ttgagtagta aaataaatag atagagaaaa 5641 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 5701 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 5761 tcacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt 5821 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg 5881 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 5941 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 6001 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 6061 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 6121 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 6181 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat 6241 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta 6301 taatacgaca aggtgaggaa ctaaaccatg accgagtaca agcccacggt gcgcctcgcc 6361 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc 6421 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa 6481 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc 6541 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc 6601 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc 6661 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg 6721 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc 6781 gagcgcgccg gggtgcccgc cttcctggag acatccgcgc cccgcaacct ccccttctac 6841 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg 6901 tgcatgaccc gcaagcccgg tgcctgacac gtgctacgag atttcgattc caccgccgcc 6961 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 7021 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 7081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 7141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 7201 tctagctaga gcttggcgta atcatggtca ttaccaatgc ttaatcagtg aggcacctat 7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 7321 tacgatacgg gagggcttac catctggccc cagcgctgcg atgataccgc gagaaccacg 7381 ctcaccggct ccggatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt 7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tattcttcct 8101 t //