LOCUS       sequence-412904-        8035 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 8035)
  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 8035)
  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-SEMA4C-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(202..221)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    897..1018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(908..927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     962..981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1148..1747)
                     /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(1231..1251)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1728..1747
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1766..1813)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1861..2190)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1875..2010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1929..1948)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2175..2195
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2313..2424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2413..2430
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2570..2587)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2588..2632)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(5378..5397)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(5410..5428)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5411..5428)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5638..5662)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(5659..5862)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5688..5708)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5863..6242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    6409..6425
                     /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(6881..6898)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6881..6897)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6996..7013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7167..7755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7247..7266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7841..7860)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(7866..7885)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        7927..8018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8019..8035,1..52,53..844)
                     /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"
ORIGIN
        1 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
       61 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      121 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      181 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
      241 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      301 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      361 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      421 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      481 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag cgatggcaac
      541 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      601 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
      661 ctggtttatt gctgataaat ccggagccgg tgagcgtggt tctcgcggta tcatcgcagc
      721 gctggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      781 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      841 gtaatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag
      901 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      961 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     1021 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     1081 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgta
     1141 gcacgtgtca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     1201 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     1261 atgtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     1321 cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc
     1381 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     1441 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     1501 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     1561 tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga
     1621 tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga
     1681 gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact
     1741 cggtcatggt ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg
     1801 attaattgtc aacacgtgct gatcagatcc gaaaatggat atacaagctc ccgggagctt
     1861 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     1921 agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     1981 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     2041 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     2101 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     2161 tggggacttt ccacacccta actgacacac attccacaga attaattcgc gttaaatttt
     2221 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2281 aaagaataga ccgagatagg gttgagtgtt gtgatgcaat ttcctcattt tattaggaaa
     2341 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     2401 atggctggca actagaaggc acagtcgagg ctgattttct ctatctattt attttactac
     2461 tcaagactat cttgcaaagt agaagaaccc tgtctgcagt atgacgaccg tgtatacccg
     2521 attggcttac cgggtaacgc gttgtgccag tgactacagg ccggccttag tggtgatggt
     2581 ggtggtgttc atgccactca atcttctgag cctcgaaaat atcattaaga ccgctgccgc
     2641 cggggctcag gctcagggac ttctgggtgt agtggttgtg cagggcctcg tgcatcacgc
     2701 tgcagctgaa cacgttgccc tgctgccacc tggacttgtc cacggtcagc ttgctgtaca
     2761 ggaagaagct gccgtcgctg tccagcactg ggggggtggt cttgtagttg ttctcgggct
     2821 ggccgttgct ctcccactcc acggcgatat cgctggggta gaagcccttc accagacagg
     2881 tcagggacac ctggttcttg gtcatctcct cccggctggg gggcagggtg tacacctggg
     2941 gctcccgtgg ctggcccttg gccttgctga ttgtcttttc gattggggca gccagggcct
     3001 tgttggagac tttgcacttg tattctttgc cgttcagcca gtcctggtgc agcacggtca
     3061 gcacggacac caccctgtag gtgctgttgt actgctcctc tctgggcttg gtcttggcgt
     3121 tgtgcacctc cacgccgtcc acgtaccagt tgaacttcac ctctgggtcc tcgtgggaca
     3181 cggccaccac cacgcaggtc acctcggggg tcctgctgat catcagggtg tccttgggct
     3241 tgggggggaa caggaacacg gaaggccctc ccagcagttc tggagctggg cagggggggc
     3301 aggtgtgggt cttgtcgccg cccggcgcgc ctccgaggtt ttccaggggt gctctagctt
     3361 ccagtgtcac gctagggcca gcaaccacag ccaccagata tccttcggcg gccagtctag
     3421 cgccttgttc ctcgctgaag cagtggtaag cgccggcatg tctaggctgt gcagccataa
     3481 ccaccagggc ctgcagtcga gcatcataca ggaaggagcc aggctgctca gctggcagat
     3541 ctctgccgcc gaatgtccat ctggcgtggg ccagattgct gctcaggtga caaggcagca
     3601 ccagatcggt gccggccacc actgtgatgt tcttaggtgt gggccgcact ttcttagagc
     3661 cccgcagatt gcagatgccg ctggtgtcgc tggtcatcac gtgctgaatc agcaggcttc
     3721 cactgtgtcc gccaacggcc acacatctgc tggtgttcac gctccaagca cagtaaggat
     3781 ctctggccag cacgcaatcg gcgcaggatc tgtacttcat gcagtcggcc acaggcagct
     3841 gaaccagctg ggatctgctg ccggcaaaca gcagcttctt agactggctc agcaccaggc
     3901 tccgcatagg ctcttggtcg aacagttgca gttcttcgat caggtgcacc caaggtccaa
     3961 gagaaacagc cttgagcagc cagccgtcgc ctgttccgat aaacagcacg gtgtaggtgg
     4021 cgccatccag tcctgtcact ctatcggcca ccaggtgggt gaagttggtg cccttcttaa
     4081 ccagcagagg tctggaccat ctggggccga cttgttcttc catcagaggg tgtttcttca
     4141 cgaagttcag gatgttgtcg gggagttcca ggctagaggt gtagccgtgc cgtctgtgcc
     4201 agttgttgat gcagctgcca ggtcttggag aaggcacggg gtctgtgtat ctgtcccact
     4261 tctgggcctc ttcgtgatac tctttgtagg ggccctcgaa cacccgctgg atctcttcca
     4321 gctgatactc gcagatggca gacaggtaca tgtcgcccca ctgggcctga aacactccga
     4381 aaaaggtggt gttgtgccag ctggtatcct gcagggtgtg catggcctgc agctggttga
     4441 aatacagctg ccaattaggg gcgctgcaag ccagtctggc cttcaggaag gtggtccact
     4501 ttctctgcag tgttctggct ccgcccatat cgcccttgca cactctagcc actctggcca
     4561 ccacctgttc agcataacag tcgctttcca cggcgcgttc gcggaagaag aagtacacct
     4621 tgtcgtcgtc gccggtgaaa gatcccacgc tctcaggcac ataggcgctg cccacaaagt
     4681 gaggctcgtt cagccagaag gccaggtact cggtcttcat gctgtggtga gggcccatat
     4741 tccgcaggat gattggctcg gtgcccagga agttgttcag ggtagcagag tacagctcgc
     4801 catcaaccag cagtccggca tggcctttgg caggatcgta aggacacttg cccttgccgt
     4861 cctcgaactc gccgtgttcc agtgtgaagg tcagcatgtt cacgtaggtg cacttaggct
     4921 ggaaggcgta ggtgccacac acgtacaggt ggctggcgtt gtagggctgc agaaaccgga
     4981 tgaagttgaa gcactcggtc tggttgttct tgcccttctg gatgcactcg gttttctttt
     5041 ccacaggggc ttcccaagag atggcgccct gcagttccag ggcttccatg ctgaaggcga
     5101 acagggcttc tctagctccc acatacagca ggccggtagg ctcggtcagt gtcagggtca
     5161 gaaagtcctg gatgccggtc tgggaaaatc tccgcacgac tgtggccagt tcgccgctag
     5221 acacggtctt tctgggcacc agattccacc aaacttcagc ggcggccgca gcgaacacca
     5281 ggcagaagat gtagctcagg gcaatgatgg tcttcatggt ggcgctagca ggcgcgtttt
     5341 cgatagaacc gaggtgcagt tggacctggg agtggacacc tgtggagaga aaggcaaagt
     5401 ggatgttatt ctatagtgtc acctaaatcg tatgtgtatg atacataagg ttatgtatta
     5461 attgtagccg cgttctaacg aagccaaggg ggtgggccta tagactctat aggcggtact
     5521 tacgtcactc ttggcacggg gaatccgcgt tccaatgcac cgttcccggc cgcggaggct
     5581 ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat
     5641 ctgacggttc actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     5701 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     5761 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     5821 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     5881 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     5941 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     6001 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     6061 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     6121 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     6181 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     6241 ccaacatggc ggtcatattg gacatgagcc aatataaatg tacatattat gatatagata
     6301 caacgtatgc aatggccaat agccaatatt gatttatgct atataaccaa tgaataatat
     6361 ggctaatggc caatattgag tcaacgccac tgtgcatgaa gctcgtaatt gttatccgct
     6421 cacaattaag ggcgacacaa aatttattct aaatgataat aaatactgat aacatcttat
     6481 agtttgtatt atattttgta ttatcgttga catgtataat tttgatatca aaaactgatt
     6541 ttccctttat tattttcgag atttattttc ttaattctct ttaacaaact agaaatattg
     6601 tatatacaaa aaatcataaa taatagatga atagtttaat tataggtgtt catcaatcga
     6661 aaaagcaacg tatcttattt aaagtgcgtt gcttttttct catttataag gttaaataat
     6721 tctcatatat caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg ctctttccct
     6781 aaactccccc cataaaaaaa cccgccgaag cgggttttta cgttatttgc ggattaacga
     6841 ttactcgtta tcagaaccgc ccagggggcc cgagcttaag actggccgtc gttttacaac
     6901 acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct gcttagtttg
     6961 atgcctggca gttccctact ctcgccttcc gcttcctcgc tcactgactc gctgcgctcg
     7021 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7081 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7141 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7201 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7261 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7321 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7381 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7441 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7501 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7561 gctacagagt tcttgaagtg gtgggctaac tacggctaca ctagaagaac agtatttggt
     7621 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7681 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7741 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7801 gacgcgcgcg taactcacgt taagggattt tggtcatgag caaggggtgt tatgagccat
     7861 attcaggtat aaatgggctc gcgataatgt tcagaattgg ttaattggtt gtaacactga
     7921 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7981 cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagtattcaa cattt
//
