LOCUS       sequence-400993-        8383 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 8383)
  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 8383)
  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-ISLR-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8383
                     /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             2222..2257
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2267..2290
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3950..3994
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3995..4021
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(4161..4178)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4167..4278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4396..4416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4401..4730
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4581..4716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4643..4662
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4778..4825
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4844..5443
                     /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(4844..4863)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5340..5360
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5573..5694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5610..5629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5664..5683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5747..6538,6539..6607))
                     /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     6370..6389
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6608..6699)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6741..6760
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6766..6785
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6871..7459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7360..7379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7613..7630
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7728..7745
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7729..7745
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8201..8217
                     /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 cctgtcctga accttgtgat tgcggcgaga agtacggctt
     1021 ccagatcgcc gattgcgcct acagagatct ggaaagcgtg ccacctggct tccccgccaa
     1081 tgtgacaaca ctgagcctga gcgccaacag actgcctgga cttcctgagg gcgccttcag
     1141 agaggttcca ctgctgcaat ctctgtggct ggcccacaac gagatcagaa cagttgctgc
     1201 tggcgccctg gcctctctgt ctcacctgaa aagcctggac ctgagccaca acctgatcag
     1261 cgacttcgct tggagcgacc tgcacaatct gtctgccctg cagctgctga agatggacag
     1321 caacgagctg accttcattc cccgggatgc cttcagaagc ctgagagccc tgagatctct
     1381 gcagctgaac cacaacaggc tgcacacact ggccgagggc accttcacac ctctgacagc
     1441 actgtcccac ctccagatca acgagaaccc cttcgactgc acctgtggca tcgtgtggct
     1501 gaaaacatgg gccctgacca ccgccgtgtc tatccccgag caggataata tcgcctgcac
     1561 aagccctcac gtgctgaagg gcacaccact gagtagactg cctccactgc cttgtagcgc
     1621 cccttctgtg cagctgagct accagccttc tcaggatggc gctgaactga ggcctggatt
     1681 tgtgctggcc ctgcactgtg atgtggatgg ccaacctgct ccacagctgc actggcacat
     1741 ccagattcct agcggcatcg tggaaatcac aagccccaac gtgggcaccg atggcagagc
     1801 attgcctgga acaccagtgg ccagcagcca gcctagattt caggcctttg ccaacggcag
     1861 cctgctgatc cctgatttcg gcaagctgga agagggcaca tacagctgtc tggccaccaa
     1921 tgagctgggc tctgccgaga gttctgtgga tgtggctctt gccacacctg gcgaaggcgg
     1981 agaagatacc ctgggcagaa gattccacgg caaggccgtg gaaggcaagg gctgttacac
     2041 cgtggacaac gaggtgcagc ctagcggccc tgaggacaac gtggtcatca tctacctgag
     2101 cagagccggc aatcctgaag ccgctgttgc tgaaggtgtc ccaggacaac tgcctcctgg
     2161 actgctgctg ctgggacaaa gcctgctgct gttcttcttc ctgacctcct ttggcgcgcc
     2221 ggaagatcaa gttgatccaa gattgattga tggcaagggc ggatctctgg aggtgctgtt
     2281 ccagggacca ggcggaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     2341 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     2401 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggta tgcagccctc
     2461 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2521 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2581 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2641 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     2701 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2761 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2821 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2881 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     2941 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     3001 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     3061 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     3121 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     3181 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     3241 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     3301 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     3361 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     3421 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3481 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3541 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3601 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3661 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3721 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3781 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3841 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3901 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggtggcggag gactgaacga
     3961 catcttcgag gctcagaaaa tcgaatggca cgaacatcac catcaccatc accatcacca
     4021 ttaaggccgg cctgtagtca ctggcacaac gcgttacccg gtaagccaat cgggtataca
     4081 cggtcgtcat actgcagaca gggttcttct actttgcaag atagtcttga gtagtaaaat
     4141 aaatagatag agaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4201 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4261 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     4321 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4381 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4501 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4561 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4621 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4681 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4741 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     4801 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     4861 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4921 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4981 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5041 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5101 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5161 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5221 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5281 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     5341 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5401 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     5461 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5521 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5701 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     5761 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5821 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     5881 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     5941 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6001 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     6061 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6121 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6181 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6241 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6301 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6361 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6421 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6481 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6541 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6601 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6661 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     6721 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     6781 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     6841 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6901 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6961 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7021 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     7081 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7141 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7201 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7261 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7321 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7381 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7441 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7501 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7561 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7621 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     7681 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     7741 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     7801 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     7861 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     7921 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     7981 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     8041 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     8101 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     8161 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     8221 cgagcttcat gcacagtggc gttgactcaa tattggccat tagccatatt attcattggt
     8281 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     8341 atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
