LOCUS       sequence-368611-        6538 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  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 6538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20- 
            20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
COMMENT     Sequence Label: pD649-HAsp-ILDR1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..184
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    302..413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     402..419
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(559..576)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(577..621)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1870..1889)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1902..1920)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1903..1920)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2130..2154)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2151..2354)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2180..2200)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2355..2734
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2901..2917
                     /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)."
     primer_bind     complement(3373..3390)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3373..3389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3488..3505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3659..4247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3739..3758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4333..4352)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4358..4377)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4419..4510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4511..4579,4580..5371)
                     /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     complement(4729..4748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5424..5545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5435..5454)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5489..5508
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5675..6274)
                     /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(5758..5778)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6255..6274
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6293..6340)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6388..6538,1..179))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6402..6537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6456..6475)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
       61 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      121 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      181 ttccacagaa ttaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
      241 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
      301 tgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
      361 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
      421 tgattttctc tatctattta ttttactact caagactatc ttgcaaagta gaagaaccct
      481 gtctgcagta tgacgaccgt gtatacccga ttggcttacc gggtaacgcg ttgtgccagt
      541 gactacaggc cggccttagt ggtgatggtg gtggtgttca tgccactcaa tcttctgagc
      601 ctcgaaaata tcattaagac cgctgccgcc ggggctcagg ctcagggact tctgggtgta
      661 gtggttgtgc agggcctcgt gcatcacgct gcagctgaac acgttgccct gctgccacct
      721 ggacttgtcc acggtcagct tgctgtacag gaagaagctg ccgtcgctgt ccagcactgg
      781 gggggtggtc ttgtagttgt tctcgggctg gccgttgctc tcccactcca cggcgatatc
      841 gctggggtag aagcccttca ccagacaggt cagggacacc tggttcttgg tcatctcctc
      901 ccggctgggg ggcagggtgt acacctgggg ctcccgtggc tggcccttgg ccttgctgat
      961 tgtcttttcg attggggcag ccagggcctt gttggagact ttgcacttgt attctttgcc
     1021 gttcagccag tcctggtgca gcacggtcag cacggacacc accctgtagg tgctgttgta
     1081 ctgctcctct ctgggcttgg tcttggcgtt gtgcacctcc acgccgtcca cgtaccagtt
     1141 gaacttcacc tctgggtcct cgtgggacac ggccaccacc acgcaggtca cctcgggggt
     1201 cctgctgatc atcagggtgt ccttgggctt gggggggaac aggaacacgg aaggccctcc
     1261 cagcagttct ggagctgggc agggggggca ggtgtgggtc ttgtcgccgc ccggcgcgcc
     1321 atgcaggacg atcagcttca cttctttgtc aggatcgccg ctggtgtcgc caggggcctc
     1381 aattgtacag tagtacacgc cgtggtccca ccacatgact tcgttgatga ccaggtcggc
     1441 ccggttctgg atggtgatct ttctctgccg gtagtcgacg cccagcacag gctcattctg
     1501 gcctcttctc tgagcaacga tccgcacttc ccgctggttg tcgttgcagt cgttgctagg
     1561 atcttggccc agagacagag cggcctgata gctggcgctg tagtagtcga agatggggtc
     1621 cttgcagaag ctcttgaatc tccaagtgac caccacatcc tgcagctggg cgcttgtggt
     1681 gtagtcgcac ttcaggatga tgctggcgaa cagggtcacg tatctctcgg tgtgctgcac
     1741 tgtaaccagc aaggcggccg cagcgaacac caggcagaag atgtagctca gggcaatgat
     1801 ggtcttcatg gtggcgctag caggcgcgtt ttcgatagaa ccgaggtgca gttggacctg
     1861 ggagtggaca cctgtggaga gaaaggcaaa gtggatgtta ttctatagtg tcacctaaat
     1921 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgaagccaag
     1981 ggggtgggcc tatagactct ataggcggta cttacgtcac tcttggcacg gggaatccgc
     2041 gttccaatgc accgttcccg gccgcggagg ctggatcggt cccggtgtct tctatggagg
     2101 tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt
     2161 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     2221 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     2281 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     2341 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     2401 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     2461 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     2521 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     2581 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     2641 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     2701 gattactatt aataactagt caataatcaa tgccaacatg gcggtcatat tggacatgag
     2761 ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca atagccaata
     2821 ttgatttatg ctatataacc aatgaataat atggctaatg gccaatattg agtcaacgcc
     2881 actgtgcatg aagctcgtaa ttgttatccg ctcacaatta agggcgacac aaaatttatt
     2941 ctaaatgata ataaatactg ataacatctt atagtttgta ttatattttg tattatcgtt
     3001 gacatgtata attttgatat caaaaactga ttttcccttt attattttcg agatttattt
     3061 tcttaattct ctttaacaaa ctagaaatat tgtatataca aaaaatcata aataatagat
     3121 gaatagttta attataggtg ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg
     3181 ttgctttttt ctcatttata aggttaaata attctcatat atcaagcaaa gtgacaggcg
     3241 cccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga
     3301 agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc gcccaggggg
     3361 cccgagctta agactggccg tcgttttaca acacagaaag agtttgtaga aacgcaaaaa
     3421 ggccatccgt caggggcctt ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt
     3481 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3541 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3601 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3661 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3721 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3781 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3841 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3901 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3961 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4021 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtgggcta
     4081 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4141 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4201 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4261 tcttttctac ggggtctgac gctcagtgga acgacgcgcg cgtaactcac gttaagggat
     4321 tttggtcatg agcaaggggt gttatgagcc atattcaggt ataaatgggc tcgcgataat
     4381 gttcagaatt ggttaattgg ttgtaacact gacccctatt tgtttatttt tctaaataca
     4441 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4501 aaggaagaat atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4561 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4621 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4681 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4741 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4801 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4861 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4921 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4981 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5041 caccacgatg cctgtagcga tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5101 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5161 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatccg gagccggtga
     5221 gcgtggttct cgcggtatca tcgcagcgct ggggccagat ggtaagccct cccgtatcgt
     5281 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5341 gataggtgcc tcactgatta agcattggta atgaccatga ttacgccaag ctctagctag
     5401 aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5461 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5521 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
     5581 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     5641 aggcggcggt ggaatcgaaa tctcgtagca cgtgtcaggc accgggcttg cgggtcatgc
     5701 accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct
     5761 cgtagaaggg gaggttgcgg ggcgcggatg tctccaggaa ggcgggcacc ccggcgcgct
     5821 cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg
     5881 gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga
     5941 ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc
     6001 cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg
     6061 cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga
     6121 gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg
     6181 cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg
     6241 tggcgaggcg caccgtgggc ttgtactcgg tcatggttta gttcctcacc ttgtcgtatt
     6301 atactatgcc gatatactat gccgatgatt aattgtcaac acgtgctgat cagatccgaa
     6361 aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     6421 tcactacttc tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa
     6481 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatg
//
