LOCUS       sequence-405796-        7663 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 7663)
  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 7663)
  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-LSR-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7663
                     /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             1502..1537
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1547..1570
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3230..3274
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3275..3301
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3441..3458)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3447..3558
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3676..3696)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3681..4010
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3861..3996
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3923..3942
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4058..4105
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4124..4723
                     /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(4124..4143)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4620..4640
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4853..4974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4890..4909)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4944..4963
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5027..5818,5819..5887))
                     /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     5650..5669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5888..5979)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6021..6040
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6046..6065
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6151..6739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6640..6659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6893..6910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7008..7025
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7009..7025
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7481..7497
                     /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 ccattcaagt gaccgtgtcc aatccttacc acgtcgtgat
     1021 cctgttccag cctgtgacac tgccctgcac ctaccagatg accagcacac ctacacagcc
     1081 catcgtgatc tggaagtaca agagcttctg ccgggacaga atcgccgatg ccttttctcc
     1141 cgcctccgtg gacaatcagc tgaatgctca gctggccgct ggcaaccctg gctacaatcc
     1201 ttatgtggaa tgccaggaca gcgtgcggac agtcagagtg gtggccacaa agcagggaaa
     1261 tgccgtgaca ctgggcgact actaccaggg cagaagaatc accatcaccg gcaacgccga
     1321 cctgaccttt gatcagacag cttggggcga cagcggcgtg tactactgtt ctgtggtgtc
     1381 tgcccaggac ctgcagggca acaatgaagc ctatgccgag ctgatcgtgc tgggcagaac
     1441 aagcggagtt gctgagctgc tgcctggatt tcaggccgga cctatcgagg atggcgcgcc
     1501 ggaagatcaa gttgatccaa gattgattga tggcaagggc ggatctctgg aggtgctgtt
     1561 ccagggacca ggcggaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     1621 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     1681 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggta tgcagccctc
     1741 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     1801 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     1861 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     1921 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     1981 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2041 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2101 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2161 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     2221 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     2281 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     2341 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2401 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2461 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2521 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     2581 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     2641 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     2701 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     2761 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     2821 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     2881 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     2941 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3001 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3061 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3121 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3181 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggtggcggag gactgaacga
     3241 catcttcgag gctcagaaaa tcgaatggca cgaacatcac catcaccatc accatcacca
     3301 ttaaggccgg cctgtagtca ctggcacaac gcgttacccg gtaagccaat cgggtataca
     3361 cggtcgtcat actgcagaca gggttcttct actttgcaag atagtcttga gtagtaaaat
     3421 aaatagatag agaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3481 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3541 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     3601 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3661 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3841 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3901 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3961 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4021 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     4081 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     4141 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4201 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4261 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4321 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4381 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4441 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4501 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4561 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     4621 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4681 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     4741 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4801 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4861 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4921 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4981 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     5041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     5161 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     5221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     5341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5881 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     6001 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     6061 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     6121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6301 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     6361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6781 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6841 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6901 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     6961 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     7021 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     7081 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     7141 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     7201 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     7261 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     7321 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     7381 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     7441 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     7501 cgagcttcat gcacagtggc gttgactcaa tattggccat tagccatatt attcattggt
     7561 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     7621 atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
