LOCUS       sequence-401001-        8158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to COMP5AP-AviTag-9xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8158)
  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 8158)
  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-SIGLEC7-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1997..2032
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2042..2065
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3725..3769
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3770..3796
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3936..3953)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3942..4053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4171..4191)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4176..4505
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4356..4491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4418..4437
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4553..4600
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4619..5218
                     /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(4619..4638)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5115..5135
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5348..5469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5385..5404)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5439..5458
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5522..6313,6314..6382))
                     /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     6145..6164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6383..6474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6516..6535
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6541..6560
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6646..7234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7135..7154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7388..7405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7503..7520
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7504..7520
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7976..7992
                     /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 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cccagaagtc caacagaaag gactacagcc tgaccatgca
     1021 gagcagcgtg accgtgcaag aggggatgtg cgttcacgtc cggtgcagct ttagctaccc
     1081 tgtggatagc cagaccgaca gcgatcctgt gcacggctat tggttcagag ccggcaacga
     1141 catctcttgg aaagcccctg tggccaccaa caatcctgcc tgggctgtgc aagaagagac
     1201 acgggacaga ttccatctgc tgggcgaccc tcagaccaag aactgtaccc tgagcatccg
     1261 ggacgccaga atgtctgatg ccggcagata cttcttccgg atggaaaagg gcaacatcaa
     1321 gtggaactat aagtacgacc agctgagcgt gaacgtgaca gccctgacac acagacccaa
     1381 cattctgatc cccggcacac tggaaagcgg ctgcttccag aatctgacct gttctgtgcc
     1441 ttgggcctgt gaacagggca cccctcctat gatcagctgg atgggcacat ccgtgtctcc
     1501 actgcaccct agcaccacca gatctagcgt gctgacactg atccctcagc ctcagcacca
     1561 cggaacaagc ctgacctgtc aagttacact gcctggcgct ggcgtgacca ccaacagaac
     1621 aatccagctc aacgtgtcct atcctcctca gaacctgacc gtgacagtgt tccaaggcga
     1681 gggaacagcc tctacagccc tgggcaatag cagcagcctg tctgtgctgg aaggccagtc
     1741 tctgagactc gtgtgcgccg tggatagcaa tcctcctgcc agactgagct ggacatggcg
     1801 gagtctgaca ctgtacccca gccagccttc taaccctctg gtgctggaac tgcaggtcca
     1861 cctgggagat gagggcgagt ttacctgcag agcccagaat agcctgggct ctcagcacgt
     1921 gtccctgaac ctgagcctgc agcaagagta cacaggcaag atgaggcctg tgtccggcgt
     1981 gttgcttggc gcgccggaag atcaagttga tccaagattg attgatggca agggcggatc
     2041 tctggaggtg ctgttccagg gaccaggcgg aagcggcaat cccggcggcg atctggcccc
     2101 ccagatgctg cgcgagctgc aggagaccaa tgccgccctg caggacgtgc gcgagctgct
     2161 gcgccaccgc gtgaaggaga tcaccttcct gaagaacacc gtgatggagt gcgacgcctg
     2221 cggtatgcag ccctctagag gctccggctc catcatccca gttgaggagg agaacccgga
     2281 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     2341 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     2401 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     2461 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     2521 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     2581 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     2641 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     2701 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     2761 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     2821 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     2881 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     2941 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     3001 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     3061 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     3121 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     3181 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     3241 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     3301 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     3361 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     3421 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     3481 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     3541 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     3601 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     3661 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtgg
     3721 cggaggactg aacgacatct tcgaggctca gaaaatcgaa tggcacgaac atcaccatca
     3781 ccatcaccat caccattaag gccggcctgt agtcactggc acaacgcgtt acccggtaag
     3841 ccaatcgggt atacacggtc gtcatactgc agacagggtt cttctacttt gcaagatagt
     3901 cttgagtagt aaaataaata gatagagaaa atcagcctcg actgtgcctt ctagttgcca
     3961 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4021 tgtcctttcc taataaaatg aggaaattgc atcacaacac tcaaccctat ctcggtctat
     4081 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4141 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4201 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4321 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4381 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4441 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4501 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     4561 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     4621 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     4681 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     4741 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     4801 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     4861 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     4921 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     4981 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5041 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     5101 gacatccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     5161 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaca
     5221 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5281 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5341 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5461 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5521 attaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5581 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5641 ccagcgctgc gatgataccg cgagaaccac gctcaccggc tccggattta tcagcaataa
     5701 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5761 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5821 acgttgttgc catcgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5881 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5941 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6001 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6061 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6121 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6181 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6241 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6301 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6361 cacggaaatg ttgaatactc atattcttcc tttttcaata ttattgaagc atttatcagg
     6421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6481 tcagtgttac aaccaattaa ccaattctga acattatcgc gagcccattt atacctgaat
     6541 atggctcata acaccccttg ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     6601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6781 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
     6841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7261 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7321 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7381 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
     7441 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     7501 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     7561 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     7621 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     7681 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     7741 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     7801 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     7861 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     7921 ataagatgtt atcagtattt attatcattt agaataaatt ttgtgtcgcc cttaattgtg
     7981 agcggataac aattacgagc ttcatgcaca gtggcgttga ctcaatattg gccattagcc
     8041 atattattca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     8101 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
