LOCUS       sequence-492990-        8002 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 8002)
  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 8002)
  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-CEACAM6-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1551..1574)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1584..1619)
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     primer_bind     complement(2595..2614)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2627..2645)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2628..2645)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2855..2879)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2876..3079)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2905..2925)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3080..3459
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3626..3642
                     /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)."
     primer_bind     complement(4098..4115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4098..4114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4213..4230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4384..4972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4464..4483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5058..5077)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5083..5102)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        5144..5235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5236..5304,5305..6096)
                     /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     complement(5454..5473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6149..6270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6160..6179)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6214..6233
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6400..6999)
                     /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(6483..6503)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6980..6999
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7018..7065)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(7113..7442)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7127..7262
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7181..7200)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7427..7447
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    7565..7676
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7665..7682
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(7822..7848)
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             complement(7849..7893)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
ORIGIN
        1 tgctcctgca cgccgtgaac caggtgcgcc tgcgggccgc gcgcgaacac cgccacgtcc
       61 tcgcctgcgt gggtctcttc gtccaggggc actgctgact gctgccgata ctcggggctc
      121 ccgctctcgc tctcggtaac atccggccgg gcgccgtcct tgagcacata gcctggaccg
      181 tttccgtata ggaggaccgt gtaggccttc ctgtcccggg ccttgccagg ggccagcccg
      241 aagatggagc tccctcgcag ggggtagcct ccgaaggaga agacgtggga gtggtcggca
      301 gtgacgaggc tcagcgtgtc ctcctcgctg gtgagctggc ccgccctctc aatggcgtcg
      361 tcgaacatga tcgtctcagt cagtgcccgg taagccctgc tttcatgatg accatggtcg
      421 atgcgaccac cctccacgaa gaggaagaag ccgcgggggt tcctgctcag caggcgcagg
      481 gcagcctctg tcatctccat cagggagggg tccagtgtgg agtctcggtg gatctcgtat
      541 ttcatgtctc caggctcaaa gagacccatg agatgggtca cagacgggtc cagggaagcc
      601 tgcatgagct cagtgcggtt ccacacatac cgggcaccct ggcgcttcgc cagccattcc
      661 tgcaccagat tcttcccgtc cagcctggtc ccaccttggc tgtagtcatc tgggtactca
      721 gggtctgggg ttcccatgcg aaacatgtac tttcggcctc cacctaggat cacgtcaatg
      781 tccatgttgg agatgagctg cgtagcgatg tcctggcacc cctcctggcg ggccgaggca
      841 ggcacgtcgg cgtccgagta ccagttgcgg ttcaccgtgt gggcgtaggt gccggctggc
      901 gaggcgtgct gcactcgtgt ggtggttacc actcccactg acttccctgc tttcttggcc
      961 cgattcatca cggagatgac ctcgttgccg cgtgtcgtgt tgcactggtt aaagcgggcg
     1021 gctgcactca agccaatggt ctggaagttg cccttgaccc cgcacaggta ggccgtggct
     1081 gtggctccac tgtctggcac atgtttgtct acattgtatg tcttggacag agccacatat
     1141 gggaagcggt ccatggccag gggtatctca ggccccagtt tgtccttctt ctgccctttt
     1201 aggatcctgg cagctgtcac cgtagacacc cccatcccat cgcccaggaa gatgatgagg
     1261 ttcttggcgg ctgtctgtgc aggctgcagc ttcttggcgg cacccagggc ctcggctgcc
     1321 tcgcggttcc agaagtccgg gttctcctcc tcaactggga tgatggagcc ggagcctcta
     1381 gagggctgca taccgcaggc gtcgcactcc atcacggtgt tcttcaggaa ggtgatctcc
     1441 ttcacgcggt ggcgcagcag ctcgcgcacg tcctgcaggg cggcattggt ctcctgcagc
     1501 tcgcgcagca tctggggggc cagatcgccg ccgggattgc cgcttccgcc tggtccctgg
     1561 aacagcacct ccagagatcc gcccttgcca tcaatcaatc ttggatcaac ttgatcttcc
     1621 ggcgcgcctg tgatcatggt cacggtggtt ctattcaggc ctgtggcgct attgtgggcc
     1681 tgacacatgt aggagccgct gttgttcacg gtgatgttgg ggatgaacag ctcttgggtg
     1741 ctctgctgga aggtgccgtt gatgaaccag gagtactgag ctggaggatt gctggcggca
     1801 tgacagctca ggttcaggtt ctcgccgggt ctgtaattgg ctttgctggg gctgattgtg
     1861 gggccgtcag ggccatacag cacattcagt gtcacggggt cgcttctatt ggcgctggca
     1921 ggattctgga tctcgcactc gtagctgccg gcatcgtttc tcttcacgct cagcagtgtc
     1981 agggtcatgt tgccgttgct cagctgcagt ctaggagaaa cgggcagaga ctggccgttg
     2041 acccaccaaa ggtaggtggt gttctgcacc tcaggctcgc aggtaaaggc cacggcgtcc
     2101 ttatcttcca cggggttgct gttgttgctg ctgatgctag gcttaggcag ctcagggtac
     2161 acgtgaaact ggccggtggc ttcctcattg accaggtcgc tcttgatcac ttgcagggtg
     2221 tagaagccgg tgtcgttctg ggtcacgttc tggatcagca ggctggcgtt agggtagatt
     2281 gtctctctgc cgctgtaggc aggtccaggt gtagcttgct gtgtgccgat cacgtagccc
     2341 acgatcaggc tattgccgtc cactctctcg cccttgtacc agctgtagcc gattctgttc
     2401 tgaggcaggt tgtgagccag cagcagcact tctttgccct cggccacgtt aaaaggggtg
     2461 ctctcgattg tcagcttggc ggccgcagcg aacaccaggc agaagatgta gctcagggca
     2521 atgatggtct tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg
     2581 acctgggagt ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc
     2641 taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag
     2701 ccaagggggt gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa
     2761 tccgcgttcc aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat
     2821 ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc
     2881 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     2941 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     3001 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     3061 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     3121 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     3181 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     3241 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     3301 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     3361 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     3421 taattgatta ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac
     3481 atgagccaat ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc
     3541 caatattgat ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca
     3601 acgccactgt gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat
     3661 ttattctaaa tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta
     3721 tcgttgacat gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt
     3781 tattttctta attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa
     3841 tagatgaata gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa
     3901 gtgcgttgct tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac
     3961 aggcgccctt aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc
     4021 gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca
     4081 gggggcccga gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc
     4141 aaaaaggcca tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc
     4201 gccttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4261 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4321 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4381 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4441 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4501 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4561 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4621 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4681 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4741 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4801 ggctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4861 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4921 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4981 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa
     5041 gggattttgg tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg
     5101 ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa
     5161 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5221 tgaaaaagga agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5341 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5401 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5521 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5641 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5761 cgtgacacca cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa
     5821 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc
     5941 ggtgagcgtg gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt
     6001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6061 gctgagatag gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta
     6121 gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca
     6181 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     6241 gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc
     6301 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
     6361 atagaaggcg gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt
     6421 catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag
     6481 ccgctcgtag aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc
     6541 gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg
     6601 gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc
     6661 caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg
     6721 cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc
     6781 caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag
     6841 gaagagttct tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg
     6901 cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc
     6961 gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc
     7021 gtattatact atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat
     7081 ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag
     7141 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
     7201 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     7261 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     7321 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     7381 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     7441 acattccaca gaattaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     7501 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     7561 ttgtgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     7621 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     7681 ggctgatttt ctctatctat ttattttact actcaagact atcttgcaaa gtagaagaac
     7741 cctgtctgca gtatgacgac cgtgtatacc cgattggctt accgggtaac gcgttgtgcc
     7801 agtgactaca ggccggcctt aatggtgatg gtgatggtga tggtgatgtt cgtgccattc
     7861 gattttctga gcctcgaaga tgtcgttcag tcctccgcca cccgggtgcg cggcgtcggt
     7921 ggtgccggcg gggggcgcca ggtcgcaggc ggtgtagggc tccaggcagg cggcgaaggc
     7981 catgacgtgc gctatgaagg tc
//
