LOCUS       sequence-385994-        8608 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 8608)
  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 8608)
  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-MERTK-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8608
                     /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             2447..2482
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2492..2515
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4175..4219
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4220..4246
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(4386..4403)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4392..4503
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4621..4641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4626..4955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4806..4941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4868..4887
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5003..5050
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5069..5668
                     /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(5069..5088)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5565..5585
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5798..5919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5835..5854)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5889..5908
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5972..6763,6764..6832))
                     /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     6595..6614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6833..6924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6966..6985
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6991..7010
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7096..7684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7585..7604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7838..7855
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7953..7970
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7954..7970
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8426..8442
                     /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 ccgcaattac agaggccaga gaggaagcca agccttatcc
     1021 tctgttcccc ggaccttttc caggcagcct gcagacagat cacacccctc tgctttctct
     1081 gcctcacgcc tctggatacc agcctgctct gatgttcagc cccacacagc ctggcagacc
     1141 tcacaccgga aatgtggcca ttcctcaagt gaccagcgtg gaaagcaagc ctctgcctcc
     1201 tctggccttt aagcacaccg tgggccacat catcctgagc gagcacaagg gcgtgaagtt
     1261 caactgcagc atcagcgtgc ccaacatcta ccaggacacc accatcagct ggtggaagga
     1321 cggaaaagaa ctgctgggag cccaccacgc catcacacag ttctaccccg atgacgaagt
     1381 gaccgccatc attgccagct tcagcatcac cagcgtgcag agaagcgaca acggcagcta
     1441 catctgcaag atgaagatca acaacgagga aatcgtcagc gaccccatct acatcgaggt
     1501 gcagggcctg cctcacttca caaagcagcc cgagagcatg aacgtgaccc ggaacaccgc
     1561 cttcaacctg acttgtcagg ctgtgggacc tcctgagcct gtgaacatct tctgggtgca
     1621 gaacagcagc agagtgaacg agcagccaga gaagtcccct agcgtgctga cagtgcctgg
     1681 actgacagag atggccgtgt ttagctgcga ggcccacaat gataagggcc tgacagtgtc
     1741 taagggcgtc cagatcaaca tcaaggccat tccatctcca cctaccgagg tgtccatccg
     1801 gaatagcaca gcccactcca tcctgatcag ctgggtgcca ggctttgacg gctacagccc
     1861 cttcagaaac tgctccatcc aagtgaaaga ggccgatcct ctgagcaacg gctccgtgat
     1921 gatcttcaac acaagcgccc tgccacacct gtaccagatt aagcagctgc aggctctggc
     1981 caactactcc atcggcgtgt cctgcatgaa cgagatcggt tggagtgccg tgtctccctg
     2041 gattctggct tctacaacag agggcgctcc tagcgtggcc cctctgaatg tgaccgtgtt
     2101 cctgaacgag agcagcgaca atgtggacat ccggtggatg aagccaccta ccaagcagca
     2161 ggatggcgaa ctcgtgggct acagaatcag ccatgtgtgg cagtccgccg gcatcagcaa
     2221 agagctgctg gaagaagtcg gccagaacgg ctccagagcc agaattagcg tccaggtgca
     2281 caacgccacc tgtacagtta gaatcgccgc cgtgacaaga ggcggcgtgg gaccattttc
     2341 cgatccagtg aagatcttta tccccgctca cggctgggtc gactacgccc catcttctac
     2401 accagctcca ggcaatgccg atcctgtgct gatcattggc gcgccggaag atcaagttga
     2461 tccaagattg attgatggca agggcggatc tctggaggtg ctgttccagg gaccaggcgg
     2521 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     2581 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     2641 gaagaacacc gtgatggagt gcgacgcctg cggtatgcag ccctctagag gctccggctc
     2701 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     2761 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     2821 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     2881 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     2941 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     3001 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     3061 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     3121 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     3181 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     3241 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     3301 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     3361 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     3421 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     3481 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     3541 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     3601 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     3661 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     3721 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     3781 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     3841 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     3901 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     3961 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     4021 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     4081 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     4141 cggcaccacc gacgccgcgc acccgggtgg cggaggactg aacgacatct tcgaggctca
     4201 gaaaatcgaa tggcacgaac atcaccatca ccatcaccat caccattaag gccggcctgt
     4261 agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc gtcatactgc
     4321 agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata gatagagaaa
     4381 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4441 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4501 atcacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4561 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4621 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4681 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4741 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4801 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4861 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4921 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4981 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     5041 ataatacgac aaggtgagga actaaaccat gaccgagtac aagcccacgg tgcgcctcgc
     5101 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5161 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5221 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5281 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5341 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5401 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5461 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5521 cgagcgcgcc ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta
     5581 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5641 gtgcatgacc cgcaagcccg gtgcctgaca cgtgctacga gatttcgatt ccaccgccgc
     5701 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5761 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5881 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5941 ctctagctag agcttggcgt aatcatggtc attaccaatg cttaatcagt gaggcaccta
     6001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6061 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     6121 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6241 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     6301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     6841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6901 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     6961 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg ctcatgacca
     7021 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
     7081 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7201 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7261 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7681 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7741 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7801 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7861 ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc tgacggatgg
     7921 cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt cttaagctcg
     7981 ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg taaaaacccg
     8041 cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag aatatttaag
     8101 ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg agaaaaaagc
     8161 aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata attaaactat
     8221 tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa agagaattaa
     8281 gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa attatacatg
     8341 tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt attatcattt
     8401 agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc ttcatgcaca
     8461 gtggcgttga ctcaatattg gccattagcc atattattca ttggttatat agcataaatc
     8521 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     8581 ggctcatgtc caatatgacc gccatgtt
//
