LOCUS       sequence-405794-        7651 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 7651)
  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 7651)
  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-ILDR2-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7651
                     /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             1490..1525
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1535..1558
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3218..3262
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3263..3289
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3429..3446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3435..3546
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3664..3684)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3669..3998
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3849..3984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3911..3930
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4046..4093
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4112..4711
                     /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(4112..4131)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4608..4628
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4841..4962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4878..4897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4932..4951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5015..5806,5807..5875))
                     /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     5638..5657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5876..5967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6009..6028
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6034..6053
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6139..6727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6628..6647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6881..6898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6996..7013
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6997..7013
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7469..7485
                     /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 ccttgcaagt gaccgtgcct gataagaaaa aggtggccat
     1021 gctgttccag cctaccgtgc tgagatgcca cttcagcaca tcttctcacc agcctgccgt
     1081 ggtgcagtgg aagttcaaga gctactgcca ggacagaatg ggcgagagcc tgggcatgag
     1141 tagcacaaga gcccagagcc tgagcaagcg gaacctggaa tgggaccctt acctggactg
     1201 tctggatagc agacggacag tcagagtggt ggccagcaag cagggatcta cagtgaccct
     1261 gggcgacttc taccggggca gagaaatcac cattgtgcac gacgccgacc tgcagatcgg
     1321 aaagctgatg tggggcgata gcggcctgta ctactgcatc atcaccacac ctgacgacct
     1381 ggaaggcaag aacgaggact ctgtggaact gctggtgctg ggcagaacag gactgctggc
     1441 tgatctgctg cctagcttcg ccgtggaaat catgcccgaa ggcgcgccgg aagatcaagt
     1501 tgatccaaga ttgattgatg gcaagggcgg atctctggag gtgctgttcc agggaccagg
     1561 cggaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac
     1621 caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt
     1681 cctgaagaac accgtgatgg agtgcgacgc ctgcggtatg cagccctcta gaggctccgg
     1741 ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     1801 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     1861 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     1921 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     1981 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     2041 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     2101 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     2161 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     2221 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     2281 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     2341 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     2401 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     2461 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     2521 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     2581 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     2641 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     2701 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     2761 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     2821 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     2881 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     2941 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     3001 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     3061 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     3121 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     3181 cgccggcacc accgacgccg cgcacccggg tggcggagga ctgaacgaca tcttcgaggc
     3241 tcagaaaatc gaatggcacg aacatcacca tcaccatcac catcaccatt aaggccggcc
     3301 tgtagtcact ggcacaacgc gttacccggt aagccaatcg ggtatacacg gtcgtcatac
     3361 tgcagacagg gttcttctac tttgcaagat agtcttgagt agtaaaataa atagatagag
     3421 aaaatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3481 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3541 tgcatcacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3601 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3661 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3721 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3841 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3901 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3961 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4021 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     4081 agtataatac gacaaggtga ggaactaaac catgaccgag tacaagccca cggtgcgcct
     4141 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4201 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     4261 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4321 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4381 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4441 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4501 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4561 ggccgagcgc gccggggtgc ccgccttcct ggagacatcc gcgccccgca acctcccctt
     4621 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4681 ctggtgcatg acccgcaagc ccggtgcctg acacgtgcta cgagatttcg attccaccgc
     4741 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4801 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4861 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4921 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4981 gacctctagc tagagcttgg cgtaatcatg gtcattacca atgcttaatc agtgaggcac
     5041 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5101 taactacgat acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac
     5161 cacgctcacc ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5221 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5281 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg
     5341 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5401 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5461 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5521 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5581 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5641 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5701 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5761 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5821 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct
     5881 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5941 ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc
     6001 tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc ttgctcatga
     6061 ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga
     6121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6301 gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6781 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6841 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6901 agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc ccctgacgga
     6961 tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc agtcttaagc
     7021 tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata acgtaaaaac
     7081 ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt cagaatattt
     7141 aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa atgagaaaaa
     7201 agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct ataattaaac
     7261 tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt taaagagaat
     7321 taagaaaata aatctcgaaa ataataaagg gaaaatcagt ttttgatatc aaaattatac
     7381 atgtcaacga taatacaaaa tataatacaa actataagat gttatcagta tttattatca
     7441 tttagaataa attttgtgtc gcccttaatt gtgagcggat aacaattacg agcttcatgc
     7501 acagtggcgt tgactcaata ttggccatta gccatattat tcattggtta tatagcataa
     7561 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     7621 attggctcat gtccaatatg accgccatgt t
//
