LOCUS       sequence-420995-        7183 bp    DNA     circular SYN 15-DEC-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 7183)
  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 7183)
  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"; 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-AMIGO2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        124..503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        504..707
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     658..678
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     704..728
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        938..956
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     938..955
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     969..988
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2882..2926
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2927..2944
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3084..3101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3090..3201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3319..3339)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3324..3653
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3504..3639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3566..3585
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3701..3748
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3767..4366
                     /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(3767..3786)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4263..4283
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4496..4617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4533..4552)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4587..4606
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4670..5461,5462..5530))
                     /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     5293..5312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5531..5622)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5664..5683
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5689..5708
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5794..6382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6283..6302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6536..6553
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6651..6668
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6652..6668
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7124..7140
                     /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)."
ORIGIN
        1 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
       61 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
      121 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      481 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      541 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      601 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      661 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      721 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      781 gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt gccaagagtg
      841 acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta gaacgcggct
      901 acaattaata cataacctta tgtatcatac acatacgatt taggtgacac tatagaataa
      961 catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc acctcggttc
     1021 tatcgaaaac gcgcctgcta gcgccaccat gaagaccatc attgccctga gctacatctt
     1081 ctgcctggtg ttcgctgcgg ccgccgtttg tccaacagcc tgtatctgtg ccaccgacat
     1141 cgtgtcctgc accaacaaga atctgagcaa ggtgcccggc aacctgttcc ggctgatcaa
     1201 gagactggac ctgagctaca accggatcgg cctgctggat agcgagtgga ttcctgtgtc
     1261 cttcgccaag ctgaacaccc tgatcctgcg gcacaacaac atcaccagca tcagcaccgg
     1321 cagcttcagc accacaccta acctgaagtg cctggacctg tccagcaaca agctgaaaac
     1381 cgtgaagaac gccgtgttcc aagagctgaa ggtgctggaa gtgctgctgc tgtacaacaa
     1441 ccacatcagc tatctggacc ctagcgcctt tggcggactg tctcagctgc agaagctgta
     1501 cctgagcggc aacttcctga cacagttccc catggacctg tacgtgggca gattcaagct
     1561 ggccgagctg atgttcctgg acgtgtccta caacagaatc cccagcatgc ccatgcacca
     1621 catcaacctg gttccaggca agcagctgag gggcatctat ctgcacggca accctttcgt
     1681 gtgcgactgc tctctgtaca gcctgctggt gttctggtac agacggcact tctccagcgt
     1741 gatggacttc aagaacgact acacctgtcg gctttggagc gacagcagac acagtagaca
     1801 ggtcctgctg ctgcaagaca gcttcatgaa ctgcagcgac tccatcatca acggaagctt
     1861 cagagccctg ggcttcatcc acgaggccca agtgggagaa agactgatgg tgcactgcga
     1921 cagcaagacc ggcaacgcca acaccgattt catctgggtc ggacccgaca acagactgct
     1981 ggaacccgac aaagagatgg aaaacttcta cgtgttccac aacggcagcc tggtcatcga
     2041 gagccctaga tttgaagatg ccggcgtgta ctcctgtatc gccatgaaca agcagcggct
     2101 gctgaacgag acagtggacg tgaccatcaa cgtgtccaac ttcaccgtgt ccagaagcca
     2161 cgctcacgag gccttcaaca caggcgcgcc gggcggcgac aagacccaca cctgcccccc
     2221 ctgcccagct ccagaactgc tgggagggcc ttccgtgttc ctgttccccc ccaagcccaa
     2281 ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg ccgtgtccca
     2341 cgaggaccca gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa
     2401 gaccaagccc agagaggagc agtacaacag cacctacagg gtggtgtccg tgctgaccgt
     2461 gctgcaccag gactggctga acggcaaaga atacaagtgc aaagtctcca acaaggccct
     2521 ggctgcccca atcgaaaaga caatcagcaa ggccaagggc cagccacggg agccccaggt
     2581 gtacaccctg ccccccagcc gggaggagat gaccaagaac caggtgtccc tgacctgtct
     2641 ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagagcaacg gccagcccga
     2701 gaacaactac aagaccaccc ccccagtgct ggacagcgac ggcagcttct tcctgtacag
     2761 caagctgacc gtggacaagt ccaggtggca gcagggcaac gtgttcagct gcagcgtgat
     2821 gcacgaggcc ctgcacaacc actacaccca gaagtccctg agcctgagcc ccggcggcag
     2881 cggtcttaat gatattttcg aggctcagaa gattgagtgg catgaacacc accaccatca
     2941 ccactaaggc cggcctgtag tcactggcac aacgcgttac ccggtaagcc aatcgggtat
     3001 acacggtcgt catactgcag acagggttct tctactttgc aagatagtct tgagtagtaa
     3061 aataaataga tagagaaaat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3121 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3181 ataaaatgag gaaattgcat cacaacactc aaccctatct cggtctattc ttttgattta
     3241 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3301 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3541 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3601 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3661 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     3721 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga ccgagtacaa
     3781 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3841 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3901 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3961 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4021 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     4081 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     4141 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     4201 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga catccgcgcc
     4261 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     4321 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg tgctacgaga
     4381 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4441 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4501 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4561 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4621 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat taccaatgct
     4681 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4741 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
     4801 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     4861 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4921 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4981 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5041 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5101 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5161 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5221 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5281 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5341 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5401 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5461 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5521 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5581 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
     5641 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
     5701 accccttgct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     5761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5941 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     6001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6421 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6481 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6541 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     6601 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     6661 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     6721 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     6781 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     6841 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     6901 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     6961 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     7021 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     7081 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     7141 ttacgagctt catgcacagt ggcgttgact caatattggc cat
//
