LOCUS       sequence-401016-        7600 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 7600)
  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 7600)
  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-PDCD1-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7600
                     /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             1439..1474
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1484..1507
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3167..3211
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3212..3238
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3378..3395)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3384..3495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3613..3633)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3618..3947
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3798..3933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3860..3879
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3995..4042
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4061..4660
                     /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(4061..4080)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4557..4577
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4790..4911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4827..4846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4881..4900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4964..5755,5756..5824))
                     /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     5587..5606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5825..5916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5958..5977
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5983..6002
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6088..6676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6577..6596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6830..6847
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6945..6962
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6946..6962
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7418..7434
                     /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 ccggatggtt tctggattct cccgatagac cctggaatcc
     1021 tcctacattc agccccgctc tgctggtggt cacagagggc gataatgcca ccttcacctg
     1081 tagcttcagc aacaccagcg agagcttcgt gctgaactgg tacagaatga gccccagcaa
     1141 ccagaccgac aagctggccg cctttcctga ggatagaagc cagcctggac aggactgccg
     1201 gttcagagtg acacagctgc ccaacggcag agacttccac atgtctgttg tgcgggccag
     1261 aagaaacgac agcggcacat atctgtgcgg cgccatttct ctggccccta aggctcagat
     1321 caaagagagc ctgagagccg agctgagagt gaccgaaaga cgggctgaag tgcccacagc
     1381 tcacccatct ccatctccaa gacctgccgg ccagtttcag accctcgttg gcgcgccgga
     1441 agatcaagtt gatccaagat tgattgatgg caagggcgga tctctggagg tgctgttcca
     1501 gggaccaggc ggaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     1561 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     1621 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggtatgc agccctctag
     1681 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     1741 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     1801 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     1861 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     1921 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     1981 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     2041 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     2101 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     2161 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     2221 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     2281 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     2341 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     2401 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     2461 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     2521 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     2581 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     2641 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     2701 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     2761 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     2821 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     2881 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     2941 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     3001 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     3061 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     3121 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt ggcggaggac tgaacgacat
     3181 cttcgaggct cagaaaatcg aatggcacga acatcaccat caccatcacc atcaccatta
     3241 aggccggcct gtagtcactg gcacaacgcg ttacccggta agccaatcgg gtatacacgg
     3301 tcgtcatact gcagacaggg ttcttctact ttgcaagata gtcttgagta gtaaaataaa
     3361 tagatagaga aaatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3421 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3481 tgaggaaatt gcatcacaac actcaaccct atctcggtct attcttttga tttataaggg
     3541 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3601 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3781 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3841 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3901 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3961 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     4021 tatcggcata gtataatacg acaaggtgag gaactaaacc atgaccgagt acaagcccac
     4081 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4141 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4201 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4261 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4321 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4381 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4441 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4501 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacatccg cgccccgcaa
     4561 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4621 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cacgtgctac gagatttcga
     4681 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4741 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4801 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4861 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4921 tataccgtcg acctctagct agagcttggc gtaatcatgg tcattaccaa tgcttaatca
     4981 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5041 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac
     5101 cgcgagaacc acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg
     5161 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5221 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta
     5281 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5341 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5401 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5461 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5521 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5581 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5641 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5701 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5761 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5821 tcatattctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5881 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt acaaccaatt
     5941 aaccaattct gaacattatc gcgagcccat ttatacctga atatggctca taacacccct
     6001 tgctcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
     6061 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6241 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
     6301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6721 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6781 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6841 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     6901 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     6961 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     7021 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     7081 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     7141 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     7201 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     7261 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     7321 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     7381 ttattatcat ttagaataaa ttttgtgtcg cccttaattg tgagcggata acaattacga
     7441 gcttcatgca cagtggcgtt gactcaatat tggccattag ccatattatt cattggttat
     7501 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     7561 tacatttata ttggctcatg tccaatatga ccgccatgtt
//
