LOCUS       sequence-401007-        7756 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 7756)
  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 7756)
  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-PDCD1LG2-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7756
                     /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             1595..1630
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1640..1663
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3323..3367
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3368..3394
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3534..3551)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3540..3651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3769..3789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3774..4103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3954..4089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4016..4035
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4151..4198
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4217..4816
                     /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(4217..4236)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4713..4733
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4946..5067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4983..5002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5037..5056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5120..5911,5912..5980))
                     /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     5743..5762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5981..6072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6114..6133
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6139..6158
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6244..6832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6733..6752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6986..7003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7101..7118
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7102..7118
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7574..7590
                     /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 ccttgtttac cgtgacagtg cccaaagagc tgtacatcat
     1021 cgagcacggc agcaacgtga ccctggaatg caatttcgat acaggctccc acgtgaacct
     1081 gggcgccatt acagcttccc tgcagaaggt ggaaaacgac acaagccctc acagagagag
     1141 agccacactg ctggaagaac agctgcctct gggcaaagcc agctttcaca tccctcaggt
     1201 gcaagtccgc gacgagggcc agtaccagtg tatcattatc tacggcgtgg cctgggacta
     1261 caagtacctg acactgaaag tgaaggccag ctaccggaag atcaacaccc acatcctgaa
     1321 ggtgcccgag acagacgagg tggaactgac ttgtcaggcc acaggctatc ctctggccga
     1381 agtgtcctgg cctaatgtgt ctgtgcctgc caacaccagc cacagcagaa caccagaggg
     1441 cctgtatcaa gtgaccagcg tgctgagact gaagcctcct ccaggcagaa acttcagctg
     1501 cgtgttctgg aacacccacg tgcgagaact gaccctggcc agcattgatc tgcagagcca
     1561 gatggaaccc cggacacacc ctacaggcgc gccggaagat caagttgatc caagattgat
     1621 tgatggcaag ggcggatctc tggaggtgct gttccaggga ccaggcggaa gcggcaatcc
     1681 cggcggcgat ctggcccccc agatgctgcg cgagctgcag gagaccaatg ccgccctgca
     1741 ggacgtgcgc gagctgctgc gccaccgcgt gaaggagatc accttcctga agaacaccgt
     1801 gatggagtgc gacgcctgcg gtatgcagcc ctctagaggc tccggctcca tcatcccagt
     1861 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     1921 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     1981 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     2041 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     2101 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     2161 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     2221 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     2281 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     2341 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     2401 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     2461 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     2521 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     2581 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     2641 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     2701 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     2761 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     2821 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     2881 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     2941 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     3001 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     3061 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     3121 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     3181 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     3241 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     3301 cgccgcgcac ccgggtggcg gaggactgaa cgacatcttc gaggctcaga aaatcgaatg
     3361 gcacgaacat caccatcacc atcaccatca ccattaaggc cggcctgtag tcactggcac
     3421 aacgcgttac ccggtaagcc aatcgggtat acacggtcgt catactgcag acagggttct
     3481 tctactttgc aagatagtct tgagtagtaa aataaataga tagagaaaat cagcctcgac
     3541 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3601 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cacaacactc
     3661 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3721 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3781 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3901 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3961 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4021 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4081 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4141 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     4201 ggtgaggaac taaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4261 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4321 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4381 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4441 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4501 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4561 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4621 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4681 ggtgcccgcc ttcctggaga catccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4741 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4801 caagcccggt gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     4861 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4921 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4981 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5041 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5101 cttggcgtaa tcatggtcat taccaatgct taatcagtga ggcacctatc tcagcgatct
     5161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5221 agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc
     5281 cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5401 cagttaatag tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt
     5461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt
     6001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6061 aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga
     6121 gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac
     6181 gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6241 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6301 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6361 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca
     6421 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6481 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6541 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6601 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6661 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6721 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6781 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6841 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6901 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6961 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaggcgagag
     7021 tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc tttttgcgtt
     7081 tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg ccccctgggc
     7141 ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct tcggcgggtt
     7201 tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg cgcctgtcac
     7261 tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa cgcactttaa
     7321 ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc atctattatt
     7381 tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga aaataaatct
     7441 cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc aacgataata
     7501 caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag aataaatttt
     7561 gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt ggcgttgact
     7621 caatattggc cattagccat attattcatt ggttatatag cataaatcaa tattggctat
     7681 tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca
     7741 atatgaccgc catgtt
//
