LOCUS       sequence-424372-        7702 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 7702)
  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 7702)
  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-FCGR1B-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7702
                     /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             1541..1576
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1586..1609
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3269..3313
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3314..3340
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3480..3497)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3486..3597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3715..3735)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3720..4049
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3900..4035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3962..3981
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4097..4144
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4163..4762
                     /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(4163..4182)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4659..4679
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4892..5013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4929..4948)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4983..5002
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5066..5857,5858..5926))
                     /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     5689..5708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5927..6018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6060..6079
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6085..6104
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6190..6778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6679..6698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6932..6949
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7047..7064
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7048..7064
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7520..7536
                     /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 cccaagtgga tacaacaaag gccgtgatca ccctgcagcc
     1021 tccatgggtg tccgtgttcc aagaggaaac cgtgacactg cactgcgagg tgctgcatct
     1081 gcctggatct agcagcaccc agtggtttct gaatggcacc gccacacaga ccagcacacc
     1141 cagctataga atcaccagcg ccagcgtgaa cgacagcggc gagtatagat gccagagagg
     1201 cctgagcggc agaagcgacc ctattcagct ggaaatccac agaggctggc tgctgctgca
     1261 ggtttccagc agagtgttca tggaaggcga gcccctggct ctgagatgtc acgcctggaa
     1321 ggacaagctg gtgtacaacg tgctgtacta ccggaacggc aaggccttca agttcttcca
     1381 ctggaacagc aacctgacca tcctcaagac caacatcagc cacaacggca cctaccactg
     1441 tagcggcatg ggcaagcaca gatacacaag cgccggcatc agccagtaca cagtgaaagg
     1501 actgcagctg cccacacctg tgtggttcca tggcgcgccg gaagatcaag ttgatccaag
     1561 attgattgat ggcaagggcg gatctctgga ggtgctgttc cagggaccag gcggaagcgg
     1621 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     1681 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     1741 caccgtgatg gagtgcgacg cctgcggtat gcagccctct agaggctccg gctccatcat
     1801 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     1861 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     1921 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     1981 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     2041 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2101 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2161 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     2221 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     2281 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     2341 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     2401 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     2461 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     2521 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     2581 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     2641 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     2701 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     2761 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     2821 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     2881 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     2941 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     3001 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3061 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3121 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3181 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     3241 caccgacgcc gcgcacccgg gtggcggagg actgaacgac atcttcgagg ctcagaaaat
     3301 cgaatggcac gaacatcacc atcaccatca ccatcaccat taaggccggc ctgtagtcac
     3361 tggcacaacg cgttacccgg taagccaatc gggtatacac ggtcgtcata ctgcagacag
     3421 ggttcttcta ctttgcaaga tagtcttgag tagtaaaata aatagataga gaaaatcagc
     3481 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3541 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca
     3601 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3661 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3721 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3841 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3901 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3961 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4021 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4081 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4141 cgacaaggtg aggaactaaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4201 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4261 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4321 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4381 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4441 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4501 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4561 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4621 cgccggggtg cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg
     4681 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4741 gacccgcaag cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta
     4801 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4861 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4921 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4981 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5041 ctagagcttg gcgtaatcat ggtcattacc aatgcttaat cagtgaggca cctatctcag
     5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5161 tacgggaggg cttaccatct ggccccagcg ctgcgatgat accgcgagaa ccacgctcac
     5221 cggctccgga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5341 gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac
     5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatattc ttcctttttc
     5941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6001 tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta
     6061 tcgcgagccc atttatacct gaatatggct cataacaccc cttgctcatg accaaaatcc
     6121 cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc
     6361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6841 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6901 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg
     6961 cgagagtagg gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt
     7021 tgcgtttcta caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc
     7081 ctgggcggtt ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg
     7141 cgggtttttt tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc
     7201 tgtcactttg cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacgca
     7261 ctttaaataa gatacgttgc tttttcgatt gatgaacacc tataattaaa ctattcatct
     7321 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
     7381 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
     7441 ataatacaaa atataataca aactataaga tgttatcagt atttattatc atttagaata
     7501 aattttgtgt cgcccttaat tgtgagcgga taacaattac gagcttcatg cacagtggcg
     7561 ttgactcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     7621 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     7681 tgtccaatat gaccgccatg tt
//
