LOCUS       sequence-401005-        8017 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 8017)
  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 8017)
  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-FCRL6-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8017
                     /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             1856..1891
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1901..1924
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3584..3628
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3629..3655
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3795..3812)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3801..3912
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4030..4050)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4035..4364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4215..4350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4277..4296
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4412..4459
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4478..5077
                     /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(4478..4497)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4974..4994
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5207..5328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5244..5263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5298..5317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5381..6172,6173..6241))
                     /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     6004..6023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6242..6333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6375..6394
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6400..6419
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6505..7093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6994..7013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7247..7264
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7362..7379
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7363..7379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7835..7851
                     /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 ccttgtatct gcaagcttgg cccaatcctg tgttcgaggg
     1021 cgacgctctg acactgagat gccaaggctg gaagaacacc cctctgagcc aagtgaagtt
     1081 ctaccgcgac ggcaagttcc tgcacttcag caaagagaat cagaccctga gcatgggagc
     1141 cgccacagtt cagagcagag gccagtactc ttgcagcggc caagtgatgt acattcccca
     1201 gaccttcaca cagaccagcg agacagccat ggtgcaggtc caagagctgt ttcctcctcc
     1261 tgtgctgtct gctatcccat ctcctgagcc tagagagggc agcctggtta ccctgaggtg
     1321 tcagacaaag ctgcaccctc tgagaagcgc cctgagactg ctgttcagct tccacaagga
     1381 tggccacaca ctgcaggaca gaggacctca tcctgagctg tgtatccctg gcgccaaaga
     1441 gggcgattct ggcctgtatt ggtgcgaagt tgctcctgaa ggcggccagg tgcagaaaca
     1501 gtctcctcag ctggaagtgc gcgtgcaggc tcctgttagt agacctgtgc tgacactgca
     1561 ccacggacct gctgatcctg ctgtgggaga tatggtgcag ctgctgtgcg aagcccagag
     1621 aggaagccct cctatcctgt acagcttcta cctggacgag aagatcgtgg gcaatcactc
     1681 tgccccttgt ggcggcacaa caagcctgct gtttccagtg aagtctgagc aggacgccgg
     1741 caattacagc tgcgaggccg agaatagcgt gtccagagag agatccgagc ctaagaagct
     1801 gagcctgaag ggctcccagg tgctgtttac acccgccagc aattggggcg cgccggaaga
     1861 tcaagttgat ccaagattga ttgatggcaa gggcggatct ctggaggtgc tgttccaggg
     1921 accaggcgga agcggcaatc ccggcggcga tctggccccc cagatgctgc gcgagctgca
     1981 ggagaccaat gccgccctgc aggacgtgcg cgagctgctg cgccaccgcg tgaaggagat
     2041 caccttcctg aagaacaccg tgatggagtg cgacgcctgc ggtatgcagc cctctagagg
     2101 ctccggctcc atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc
     2161 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat
     2221 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg
     2281 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt
     2341 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac
     2401 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg
     2461 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa
     2521 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc
     2581 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc
     2641 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga
     2701 cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta
     2761 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg
     2821 gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc
     2881 cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga
     2941 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg
     3001 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca
     3061 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc
     3121 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga
     3181 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct
     3241 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc
     3301 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc
     3361 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt
     3421 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt
     3481 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc
     3541 gccccccgcc ggcaccaccg acgccgcgca cccgggtggc ggaggactga acgacatctt
     3601 cgaggctcag aaaatcgaat ggcacgaaca tcaccatcac catcaccatc accattaagg
     3661 ccggcctgta gtcactggca caacgcgtta cccggtaagc caatcgggta tacacggtcg
     3721 tcatactgca gacagggttc ttctactttg caagatagtc ttgagtagta aaataaatag
     3781 atagagaaaa tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3841 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3901 ggaaattgca tcacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3961 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4021 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4141 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4201 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4261 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4321 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4381 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     4441 cggcatagta taatacgaca aggtgaggaa ctaaaccatg accgagtaca agcccacggt
     4501 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4561 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4621 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4681 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4741 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4801 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4861 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4921 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acatccgcgc cccgcaacct
     4981 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5041 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac gtgctacgag atttcgattc
     5101 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5161 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5221 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5281 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5341 accgtcgacc tctagctaga gcttggcgta atcatggtca ttaccaatgc ttaatcagtg
     5401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5461 tgtagataac tacgatacgg gagggcttac catctggccc cagcgctgcg atgataccgc
     5521 gagaaccacg ctcaccggct ccggatttat cagcaataaa ccagccagcc ggaagggccg
     5581 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5641 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc atcgctacag
     5701 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5761 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5821 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5881 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5941 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6001 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6061 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6121 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6181 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6241 tattcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6301 acatatttga atgtatttag aaaaataaac aaataggggt cagtgttaca accaattaac
     6361 caattctgaa cattatcgcg agcccattta tacctgaata tggctcataa caccccttgc
     6421 tcatgaccaa aatcccttaa cgtgagttac gcgcgcgtcg ttccactgag cgtcagaccc
     6481 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6541 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6601 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6661 gtagccgtag ttagcccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6721 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6781 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6841 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6901 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6961 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7021 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7081 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7141 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7201 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7261 cgaggaagcg gaaggcgaga gtagggaact gccaggcatc aaactaagca gaaggcccct
     7321 gacggatggc ctttttgcgt ttctacaaac tctttctgtg ttgtaaaacg acggccagtc
     7381 ttaagctcgg gccccctggg cggttctgat aacgagtaat cgttaatccg caaataacgt
     7441 aaaaacccgc ttcggcgggt ttttttatgg ggggagttta gggaaagagc atttgtcaga
     7501 atatttaagg gcgcctgtca ctttgcttga tatatgagaa ttatttaacc ttataaatga
     7561 gaaaaaagca acgcacttta aataagatac gttgcttttt cgattgatga acacctataa
     7621 ttaaactatt catctattat ttatgatttt ttgtatatac aatatttcta gtttgttaaa
     7681 gagaattaag aaaataaatc tcgaaaataa taaagggaaa atcagttttt gatatcaaaa
     7741 ttatacatgt caacgataat acaaaatata atacaaacta taagatgtta tcagtattta
     7801 ttatcattta gaataaattt tgtgtcgccc ttaattgtga gcggataaca attacgagct
     7861 tcatgcacag tggcgttgac tcaatattgg ccattagcca tattattcat tggttatata
     7921 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     7981 atttatattg gctcatgtcc aatatgaccg ccatgtt
//
