LOCUS       sequence-426451-        7642 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 7642)
  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 7642)
  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-CD300A-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7642
                     /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             1481..1516
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1526..1549
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3209..3253
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3254..3280
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3420..3437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3426..3537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3655..3675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3660..3989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3840..3975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3902..3921
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4037..4084
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4103..4702
                     /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(4103..4122)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4599..4619
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4832..4953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4869..4888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4923..4942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5006..5797,5798..5866))
                     /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     5629..5648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5867..5958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6000..6019
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6025..6044
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6130..6718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6619..6638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6872..6889
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6987..7004
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6988..7004
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7460..7476
                     /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 ccttgagcaa gtgtagaaca gtggctggcc ctgtcggcgg
     1021 atctctgtct gttcagtgcc cctacgagaa agagcacaga accctgaaca agtactggtg
     1081 ccggcctcct cagatcttcc tgtgcgacaa gatcgtggaa acaaagggca gcgccggcaa
     1141 gagaaacggc agagtgtcta tcagagacag ccccgccaac ctgagcttca ccgtgacact
     1201 ggaaaacctg accgaggaag atgccggcac atattggtgc ggagtggaca ccccttggct
     1261 gagagacttt cacgaccccg tggtggaagt ggaagtgtcc gtgtttcctg ccagcaccag
     1321 catgacacca gcctctatta cagccgccaa gaccagcacc atcaccaccg cttttccacc
     1381 tgtgtccagc accacactgt ttgccgtggg agccacacac agcgccagca tccaagaaga
     1441 gacagaggaa gtcgtgaaca gccagctgcc tggcgcgccg gaagatcaag ttgatccaag
     1501 attgattgat ggcaagggcg gatctctgga ggtgctgttc cagggaccag gcggaagcgg
     1561 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     1621 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     1681 caccgtgatg gagtgcgacg cctgcggtat gcagccctct agaggctccg gctccatcat
     1741 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     1801 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     1861 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     1921 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     1981 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2041 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2101 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     2161 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     2221 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     2281 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     2341 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     2401 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     2461 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     2521 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     2581 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     2641 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     2701 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     2761 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     2821 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     2881 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     2941 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3001 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3061 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3121 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     3181 caccgacgcc gcgcacccgg gtggcggagg actgaacgac atcttcgagg ctcagaaaat
     3241 cgaatggcac gaacatcacc atcaccatca ccatcaccat taaggccggc ctgtagtcac
     3301 tggcacaacg cgttacccgg taagccaatc gggtatacac ggtcgtcata ctgcagacag
     3361 ggttcttcta ctttgcaaga tagtcttgag tagtaaaata aatagataga gaaaatcagc
     3421 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3481 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca
     3541 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3661 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3721 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3841 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3901 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3961 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4021 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4081 cgacaaggtg aggaactaaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4141 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4201 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4261 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4321 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4381 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4441 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4501 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4561 cgccggggtg cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg
     4621 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4681 gacccgcaag cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta
     4741 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4801 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4861 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4921 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4981 ctagagcttg gcgtaatcat ggtcattacc aatgcttaat cagtgaggca cctatctcag
     5041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5101 tacgggaggg cttaccatct ggccccagcg ctgcgatgat accgcgagaa ccacgctcac
     5161 cggctccgga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5281 gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac
     5341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatattc ttcctttttc
     5881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5941 tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta
     6001 tcgcgagccc atttatacct gaatatggct cataacaccc cttgctcatg accaaaatcc
     6061 cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6241 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc
     6301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6841 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg
     6901 cgagagtagg gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt
     6961 tgcgtttcta caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc
     7021 ctgggcggtt ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg
     7081 cgggtttttt tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc
     7141 tgtcactttg cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacgca
     7201 ctttaaataa gatacgttgc tttttcgatt gatgaacacc tataattaaa ctattcatct
     7261 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
     7321 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
     7381 ataatacaaa atataataca aactataaga tgttatcagt atttattatc atttagaata
     7441 aattttgtgt cgcccttaat tgtgagcgga taacaattac gagcttcatg cacagtggcg
     7501 ttgactcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     7561 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     7621 tgtccaatat gaccgccatg tt
//
