LOCUS       sequence-457774-        8602 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 8602)
  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 8602)
  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-KIRREL1-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1535..1554)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1567..1585)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1568..1585)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1795..1819)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1816..2019)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1845..1865)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2020..2399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2566..2582
                     /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(3038..3055)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3038..3054)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3153..3170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3324..3912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3404..3423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3998..4017)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4023..4042)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4084..4175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4176..4244,4245..5036)
                     /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(4394..4413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5089..5210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5100..5119)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5154..5173
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5340..5939)
                     /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(5423..5443)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5920..5939
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5958..6005)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6053..6382)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6067..6202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6121..6140)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6367..6387
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    6505..6616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6605..6622
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6762..6788)
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             complement(6789..6833)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(8493..8516)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(8526..8561)
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
ORIGIN
        1 cttccagctg aatgatggct gtgccaggtc caaagctgtt ccaggcggtg cagttgtagt
       61 gggtctgaaa gtcggcttcc atcacgttgt tgatggtcag ggtagacagc acgccgctgc
      121 cgctgtttgt tctttccacg gtgtaccgtt ccagggtgcc cacttccagg aagttttctt
      181 tccaggccca ggcaattctg tcaggtggag gggtgctgcc gatgaagcat tccacctttc
      241 cgccatctcc gcgaacggca tactgcacgg cctcgctgct gatgatagga gggccgttaa
      301 cgtacagggg cacttctctc tcggccacgc cgattcttgg cacgatggct ctacatgtgt
      361 aggtgccggc atcggcctgg gtcacagact tcaggagcag ctgattgctg ttgctcagca
      421 ccatgttgct gtctttcttg gtccaggtca gtgtcagagg agggtttccg acccacacac
      481 aggtcagggt cacatcagag ccgatgtcgg tggtggtagg cttagggtcc accacgattc
      541 taggggcgaa gtgcacgttg accagtgtgg acacgttggt gctgcccact ttgttgtgca
      601 cctcgcagga cacaggctcg gtgaagaagc tgtagtccac gtttgtctcg tatctgctct
      661 cgtgggcgtc ctcaatcagg aagccgcctt tggcccatct gtagcccagg atctcaggat
      721 tggcggtggc ttggcaggta aacacaactc gctcgccctc ttgcacagtc tggggctcga
      781 tggacagtgt cacggttgga ggatggtgca cgtccagctc gatgctggtt tctttgccgc
      841 taggaatggc ctcgttcatg ctccgacagg tgaacacccg gccgatgtcc agatcggtgg
      901 ggttgatcag cagctggctg acggttgtct ctctcttgcc atctttcagc agctctgtag
      961 aggccacggc gccctcttgc tgtgtgccat ctctgaacca gatgatggtg gcggcaggtt
     1021 tggcgttgaa ggctctgcag gtcaggttgt gaggtgttcc ggcctgcagc agaatcacag
     1081 ggccgccatc gattctggtg tcctcaggag ggatcagcac ggtcagcttg gcccttctac
     1141 ttctcagagc ggcctctgtg gcctgacact cataggaggc atcgtcgctc agctcggcat
     1201 cggtgatttc caggttgtac tggccggcat cagcagagcc cacaactcta taccgaggcc
     1261 aggctttcag tccctgtccc attccgagag ccagtccgtc cttggtccac tgcacgatgc
     1321 cagagtagtt cagcagcacg caaggcagca cagctctttg tccagccacc actgtctggt
     1381 cggcgggctc ttgagaaaat ctggtctgtg ttccctgggc ggccgcagcg aacaccaggc
     1441 agaagatgta gctcagggca atgatggtct tcatggtggc gctagcaggc gcgttttcga
     1501 tagaaccgag gtgcagttgg acctgggagt ggacacctgt ggagagaaag gcaaagtgga
     1561 tgttattcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt
     1621 gtagccgcgt tctaacgaag ccaagggggt gggcctatag actctatagg cggtacttac
     1681 gtcactcttg gcacggggaa tccgcgttcc aatgcaccgt tcccggccgc ggaggctgga
     1741 tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg
     1801 acggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     1861 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     1921 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     1981 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     2041 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     2101 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     2161 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     2221 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     2281 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     2341 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgcca
     2401 acatggcggt catattggac atgagccaat ataaatgtac atattatgat atagatacaa
     2461 cgtatgcaat ggccaatagc caatattgat ttatgctata taaccaatga ataatatggc
     2521 taatggccaa tattgagtca acgccactgt gcatgaagct cgtaattgtt atccgctcac
     2581 aattaagggc gacacaaaat ttattctaaa tgataataaa tactgataac atcttatagt
     2641 ttgtattata ttttgtatta tcgttgacat gtataatttt gatatcaaaa actgattttc
     2701 cctttattat tttcgagatt tattttctta attctcttta acaaactaga aatattgtat
     2761 atacaaaaaa tcataaataa tagatgaata gtttaattat aggtgttcat caatcgaaaa
     2821 agcaacgtat cttatttaaa gtgcgttgct tttttctcat ttataaggtt aaataattct
     2881 catatatcaa gcaaagtgac aggcgccctt aaatattctg acaaatgctc tttccctaaa
     2941 ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga ttaacgatta
     3001 ctcgttatca gaaccgccca gggggcccga gcttaagact ggccgtcgtt ttacaacaca
     3061 gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct tagtttgatg
     3121 cctggcagtt ccctactctc gccttccgct tcctcgctca ctgactcgct gcgctcggtc
     3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3721 acagagttct tgaagtggtg ggctaactac ggctacacta gaagaacagt atttggtatc
     3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac
     3961 gcgcgcgtaa ctcacgttaa gggattttgg tcatgagcaa ggggtgttat gagccatatt
     4021 caggtataaa tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc
     4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4141 gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat ttccgtgtcg
     4201 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4261 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4321 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4381 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4441 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4501 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4561 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4621 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4681 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcgatggca acaacgttgc
     4741 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4861 ttgctgataa atccggagcc ggtgagcgtg gttctcgcgg tatcatcgca gcgctggggc
     4921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaatgac
     5041 catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg
     5101 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5161 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg
     5221 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     5281 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tagcacgtgt
     5341 caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg
     5401 gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc ggatgtctcc
     5461 aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac gacggcgctg
     5521 cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc
     5581 tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc cagccgggaa
     5641 ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc
     5701 tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac cttgccgatg
     5761 tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc gatgtggcgg
     5821 tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt
     5881 acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg
     5941 gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga tgattaattg
     6001 tcaacacgtg ctgatcagat ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa
     6061 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gcagaggcgg
     6121 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     6181 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     6241 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     6301 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     6361 ttccacaccc taactgacac acattccaca gaattaattc gcgttaaatt tttgttaaat
     6421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6481 gaccgagata gggttgagtg ttgtgatgca atttcctcat tttattagga aaggacagtg
     6541 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
     6601 caactagaag gcacagtcga ggctgatttt ctctatctat ttattttact actcaagact
     6661 atcttgcaaa gtagaagaac cctgtctgca gtatgacgac cgtgtatacc cgattggctt
     6721 accgggtaac gcgttgtgcc agtgactaca ggccggcctt aatggtgatg gtgatggtga
     6781 tggtgatgtt cgtgccattc gattttctga gcctcgaaga tgtcgttcag tcctccgcca
     6841 cccgggtgcg cggcgtcggt ggtgccggcg gggggcgcca ggtcgcaggc ggtgtagggc
     6901 tccaggcagg cggcgaaggc catgacgtgc gctatgaagg tctgctcctg cacgccgtga
     6961 accaggtgcg cctgcgggcc gcgcgcgaac accgccacgt cctcgcctgc gtgggtctct
     7021 tcgtccaggg gcactgctga ctgctgccga tactcggggc tcccgctctc gctctcggta
     7081 acatccggcc gggcgccgtc cttgagcaca tagcctggac cgtttccgta taggaggacc
     7141 gtgtaggcct tcctgtcccg ggccttgcca ggggccagcc cgaagatgga gctccctcgc
     7201 agggggtagc ctccgaagga gaagacgtgg gagtggtcgg cagtgacgag gctcagcgtg
     7261 tcctcctcgc tggtgagctg gcccgccctc tcaatggcgt cgtcgaacat gatcgtctca
     7321 gtcagtgccc ggtaagccct gctttcatga tgaccatggt cgatgcgacc accctccacg
     7381 aagaggaaga agccgcgggg gttcctgctc agcaggcgca gggcagcctc tgtcatctcc
     7441 atcagggagg ggtccagtgt ggagtctcgg tggatctcgt atttcatgtc tccaggctca
     7501 aagagaccca tgagatgggt cacagacggg tccagggaag cctgcatgag ctcagtgcgg
     7561 ttccacacat accgggcacc ctggcgcttc gccagccatt cctgcaccag attcttcccg
     7621 tccagcctgg tcccaccttg gctgtagtca tctgggtact cagggtctgg ggttcccatg
     7681 cgaaacatgt actttcggcc tccacctagg atcacgtcaa tgtccatgtt ggagatgagc
     7741 tgcgtagcga tgtcctggca cccctcctgg cgggccgagg caggcacgtc ggcgtccgag
     7801 taccagttgc ggttcaccgt gtgggcgtag gtgccggctg gcgaggcgtg ctgcactcgt
     7861 gtggtggtta ccactcccac tgacttccct gctttcttgg cccgattcat cacggagatg
     7921 acctcgttgc cgcgtgtcgt gttgcactgg ttaaagcggg cggctgcact caagccaatg
     7981 gtctggaagt tgcccttgac cccgcacagg taggccgtgg ctgtggctcc actgtctggc
     8041 acatgtttgt ctacattgta tgtcttggac agagccacat atgggaagcg gtccatggcc
     8101 aggggtatct caggccccag tttgtccttc ttctgccctt ttaggatcct ggcagctgtc
     8161 accgtagaca cccccatccc atcgcccagg aagatgatga ggttcttggc ggctgtctgt
     8221 gcaggctgca gcttcttggc ggcacccagg gcctcggctg cctcgcggtt ccagaagtcc
     8281 gggttctcct cctcaactgg gatgatggag ccggagcctc tagagggctg cataccgcag
     8341 gcgtcgcact ccatcacggt gttcttcagg aaggtgatct ccttcacgcg gtggcgcagc
     8401 agctcgcgca cgtcctgcag ggcggcattg gtctcctgca gctcgcgcag catctggggg
     8461 gccagatcgc cgccgggatt gccgcttccg cctggtccct ggaacagcac ctccagagat
     8521 ccgcccttgc catcaatcaa tcttggatca acttgatctt ccggcgcgcc agcaatgatg
     8581 cccacaggca gcacctcgcg tt
//
