LOCUS       sequence-303979-        8521 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 8521)
  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 8521)
  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-ICAM3-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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)."
     enhancer        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4126..4161
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             4171..4194
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5854..5898
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5899..5925
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(6065..6082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6071..6182
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6300..6320)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6305..6634
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6485..6620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6547..6566
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6682..6729
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6748..7347
                     /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(6748..6767)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7244..7264
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7477..7598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7514..7533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7568..7587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7651..8442,8443..8511))
                     /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     8274..8293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8512..8521,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgccca agagtttctg
     2761 ctgagagtgg aaccccagaa tcctgtgctg tctgccggcg gaagcctgtt cgtgaattgc
     2821 agcacagatt gccccagcag cgagaagatc gccctggaaa ccagcctgag caaagaactg
     2881 gtggcctctg gaatgggctg ggccgccttt aatctgagca acgtgaccgg caactcccgc
     2941 atcctgtgca gcgtgtactg taacggaagc cagatcaccg gcagcagcaa catcaccgtg
     3001 tacagactgc ctgagagagt cgaactggct cctctgcctc cttggcagcc tgtgggacag
     3061 aatttcaccc tgcggtgtca ggtggaagat ggcagcccta gaacaagcct gaccgtggtg
     3121 ctgctgagat gggaagaaga actgagcaga cagcccgccg tggaagaacc tgctgaagtg
     3181 acagctacag tgctggcctc cagagatgat catggcgccc ctttcagctg tcggaccgaa
     3241 ctggatatgc agcctcaagg cctgggcctg tttgtgaaca catctgcccc tcggcagctg
     3301 agaaccttcg tgcttcctgt gacacctcct aggctggtgg cccctagatt cctggaagtg
     3361 gaaacctcct ggcctgtgga ctgtaccctg gatggactgt ttcctgccag cgaggcccaa
     3421 gtttatctgg ccctgggcga tcagatgctg aacgccacag tgatgaacca cggcgataca
     3481 ctgacagcca ccgctacagc cacagccaga gctgatcaag agggcgccag agaaatcgtg
     3541 tgcaatgtga cactcggcgg cgagagaaga gaggccaggg aaaatctgac cgtgttcagc
     3601 tttctgggcc ccatcgtgaa tctgagcgag cctacagctc acgagggcag cacagtgacc
     3661 gtgtcttgta tggctggcgc tagagtgcaa gtgactctgg atggtgttcc tgccgctgct
     3721 cctggacaac ctgctcagct gcaactgaat gccaccgagt ccgacgacgg cagaagcttt
     3781 ttttgctccg ccacactgga agtcgacggc gagttcctgc acagaaatag cagcgtgcag
     3841 ctgcgggtgc tgtacggccc taagatcgat agagccacct gtcctcagca cctgaagtgg
     3901 aaggacaaga ccagacacgt gctgcagtgc caggccagag gcaatcctta tcctgagctg
     3961 agatgcctga aagagggcag ctccagagaa gtgcctgtgg gcatcccttt cttcgtgaac
     4021 gtgacccaca acggcaccta ccagtgtcag gcctcaagca gcagaggcaa gtacaccctg
     4081 gtggtggtca tggatatcga ggccggcagc tctcatggcg cgccggaaga tcaagttgat
     4141 ccaagattga ttgatggcaa gggcggatct ctggaggtgc tgttccaggg accaggcgga
     4201 agcggcaatc ccggcggcga tctggccccc cagatgctgc gcgagctgca ggagaccaat
     4261 gccgccctgc aggacgtgcg cgagctgctg cgccaccgcg tgaaggagat caccttcctg
     4321 aagaacaccg tgatggagtg cgacgcctgc ggtatgcagc cctctagagg ctccggctcc
     4381 atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc cgaggccctg
     4441 ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat catcttcctg
     4501 ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg gcagaagaag
     4561 gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt ggctctgtcc
     4621 aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac ggcctacctg
     4681 tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg ctttaaccag
     4741 tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg
     4801 aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc cggcacctac
     4861 gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc ggcccgccag
     4921 gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga cgtgatccta
     4981 ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta cccagatgac
     5041 tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg gctggcgaag
     5101 cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc cctggacccg
     5161 tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga gatccaccga
     5221 gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg cctgctgagc
     5281 aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca tggtcatcat
     5341 gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc cattgagagg
     5401 gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga ccactcccac
     5461 gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct ggcccctggc
     5521 aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc aggctatgtg
     5581 ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc cgagtatcgg
     5641 cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt ggcggtgttc
     5701 gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt catagcgcac
     5761 gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc gccccccgcc
     5821 ggcaccaccg acgccgcgca cccgggtggc ggaggactga acgacatctt cgaggctcag
     5881 aaaatcgaat ggcacgaaca tcaccatcac catcaccatc accattaagg ccggcctgta
     5941 gtcactggca caacgcgtta cccggtaagc caatcgggta tacacggtcg tcatactgca
     6001 gacagggttc ttctactttg caagatagtc ttgagtagta aaataaatag atagagaaaa
     6061 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6121 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6181 tcacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6241 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     6301 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6361 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     6421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     6481 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6541 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     6601 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     6661 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     6721 taatacgaca aggtgaggaa ctaaaccatg accgagtaca agcccacggt gcgcctcgcc
     6781 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     6841 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     6901 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6961 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     7021 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     7081 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     7141 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     7201 gagcgcgccg gggtgcccgc cttcctggag acatccgcgc cccgcaacct ccccttctac
     7261 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     7321 tgcatgaccc gcaagcccgg tgcctgacac gtgctacgag atttcgattc caccgccgcc
     7381 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     7441 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     7501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7561 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     7621 tctagctaga gcttggcgta atcatggtca ttaccaatgc ttaatcagtg aggcacctat
     7681 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7741 tacgatacgg gagggcttac catctggccc cagcgctgcg atgataccgc gagaaccacg
     7801 ctcaccggct ccggatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7861 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7921 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt
     7981 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8041 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8101 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8161 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8221 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8281 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8341 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8401 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8461 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tattcttcct
     8521 t
//
