LOCUS       sequence-303831-        8215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            COMP5AP-AviTag-9xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8215)
  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 8215)
  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-CEACAM18-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8215
                     /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             3820..3855
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             3865..3888
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5548..5592
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5593..5619
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(5759..5776)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5765..5876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5994..6014)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5999..6328
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6179..6314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6241..6260
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6376..6423
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6442..7041
                     /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(6442..6461)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6938..6958
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7171..7292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7208..7227)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7262..7281
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7345..8136,8137..8205))
                     /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     7968..7987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8206..8215,1..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 gatctttatc
     2761 acacagaccc tgggcatcaa gggctacaga acagtggtgg ccctggacaa ggtgcccgag
     2821 gatgtgcaag agtacagctg gtactggggc gccaatgaca gcgccggcaa catgatcatc
     2881 agccacaagc ctccaagcgc tcagcagccc ggacctatgt acacaggccg cgagagagtg
     2941 aatagagagg gcagcctgct gatcagaccc accgctctga acgacaccgg caactacaca
     3001 gttagagtgg tggctggcaa cgagacacag agagctacag gctggctgga agtgctggaa
     3061 ctgggcagca atctgggcat ctctgtgaat gccagcagcc tggtggaaaa catggatagc
     3121 gtggccgccg actgcctgac caacgtgacc aatatcactt ggtacgtgaa cgacgtgccc
     3181 accagcagca gcgacagaat gacaatcagc cccgatggca agaccctcgt gatcctgaga
     3241 gtgtccagat acgaccggac catccagtgc atgatcgaga gcttccccga gatcttccag
     3301 cggagcgaga gaatcagcct gacagtggcc tacggacccg actacgtgct gctgagaagc
     3361 aaccccgacg acttcaatgg catcgtgaca gccgagatcg gctcccaggt ggaaatggaa
     3421 tgcatctgct acagctttct ggacctgaag taccactgga ttcacaacgg ctccctgctg
     3481 aacttcagcg acgccaagat gaatctgtcc agcctggcct gggagcagat gggcagatac
     3541 agatgcaccg tggaaaaccc cgtgacacag ctgatcatgt atatggacgt gcggattcag
     3601 gcccctcacg aggatctgac ctgctgtcct cagagattcc tgcacctgag aatccacggc
     3661 gacgtgccac atcatgccga ttctgctgga tggcggctgc atctttggag ccctgatcct
     3721 tgtagcgacc agcctctgct gaaccaggat aagagcggca gcatgagcgt gcaccccaga
     3781 cctgaggata agaccagaag ggccagcaga ggcgcgccgg aagatcaagt tgatccaaga
     3841 ttgattgatg gcaagggcgg atctctggag gtgctgttcc agggaccagg cggaagcggc
     3901 aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac caatgccgcc
     3961 ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt cctgaagaac
     4021 accgtgatgg agtgcgacgc ctgcggtatg cagccctcta gaggctccgg ctccatcatc
     4081 ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc
     4141 gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt cctgggcgat
     4201 gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa gaaggacaaa
     4261 ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct gtccaagaca
     4321 tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta cctgtgcggg
     4381 gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa ccagtgcaac
     4441 acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc agggaagtca
     4501 gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac ctacgcccac
     4561 acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg
     4621 tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat cctaggtgga
     4681 ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga tgactacagc
     4741 caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc gaagcgccag
     4801 ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga cccgtctgtg
     4861 acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca ccgagactcc
     4921 acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct gagcaggaac
     4981 ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca tcatgaaagc
     5041 agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga gagggcgggc
     5101 cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc
     5161 tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc tggcaaggcc
     5221 cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta tgtgctcaag
     5281 gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta tcggcagcag
     5341 tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt gttcgcgcgc
     5401 ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc gcacgtcatg
     5461 gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc cgccggcacc
     5521 accgacgccg cgcacccggg tggcggagga ctgaacgaca tcttcgaggc tcagaaaatc
     5581 gaatggcacg aacatcacca tcaccatcac catcaccatt aaggccggcc tgtagtcact
     5641 ggcacaacgc gttacccggt aagccaatcg ggtatacacg gtcgtcatac tgcagacagg
     5701 gttcttctac tttgcaagat agtcttgagt agtaaaataa atagatagag aaaatcagcc
     5761 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     5821 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcacaa
     5881 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5941 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     6001 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6061 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6121 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6181 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6241 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     6301 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     6361 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     6421 gacaaggtga ggaactaaac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     6481 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     6541 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     6601 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     6661 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     6721 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     6781 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     6841 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     6901 gccggggtgc ccgccttcct ggagacatcc gcgccccgca acctcccctt ctacgagcgg
     6961 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     7021 acccgcaagc ccggtgcctg acacgtgcta cgagatttcg attccaccgc cgccttctat
     7081 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     7141 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     7201 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7261 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     7321 tagagcttgg cgtaatcatg gtcattacca atgcttaatc agtgaggcac ctatctcagc
     7381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7441 acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac cacgctcacc
     7501 ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7621 ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg tggtgtcacg
     7681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct tcctt
//
