LOCUS       sequence-401019-        7528 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 7528)
  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 7528)
  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-MOG-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7528
                     /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"
     regulatory      1307..1316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1367..1402
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1412..1435
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3095..3139
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3140..3166
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3306..3323)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3312..3423
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3541..3561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3546..3875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3726..3861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3788..3807
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3923..3970
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3989..4588
                     /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(3989..4008)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4485..4505
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4718..4839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4755..4774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4809..4828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4892..5683,5684..5752))
                     /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     5515..5534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5753..5844)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5886..5905
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5911..5930
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6016..6604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6505..6524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6758..6775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6873..6890
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6874..6890
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7346..7362
                     /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 ccggacagtt tagagtgatt ggccctagac accccatcag
     1021 agccctcgtg ggagatgagg tggaactgcc ttgtagaatc agccccggca agaacgccac
     1081 aggcatggaa gttggctggt acagacctcc attcagcaga gtggtgcacc tgtacagaaa
     1141 cggcaaggac caggatggcg atcaggcccc tgagtacaga ggaagaaccg agctgctgaa
     1201 ggatgccatc ggcgagggca aagtgaccct gagaatcaga aacgtgcggt tcagcgacga
     1261 aggcggcttc acctgtttct tccgggacca cagctatcaa gaggaagccg ccatggaact
     1321 gaaggtggaa gatcccttct actgggtgtc acccggtggc gcgccggaag atcaagttga
     1381 tccaagattg attgatggca agggcggatc tctggaggtg ctgttccagg gaccaggcgg
     1441 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     1501 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     1561 gaagaacacc gtgatggagt gcgacgcctg cggtatgcag ccctctagag gctccggctc
     1621 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     1681 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     1741 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     1801 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     1861 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     1921 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     1981 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     2041 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     2101 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     2161 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     2221 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     2281 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     2341 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     2401 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     2461 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     2521 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     2581 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     2641 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     2701 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     2761 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     2821 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     2881 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     2941 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     3001 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     3061 cggcaccacc gacgccgcgc acccgggtgg cggaggactg aacgacatct tcgaggctca
     3121 gaaaatcgaa tggcacgaac atcaccatca ccatcaccat caccattaag gccggcctgt
     3181 agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc gtcatactgc
     3241 agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata gatagagaaa
     3301 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3361 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3421 atcacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3481 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3541 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3601 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3721 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3781 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3841 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3901 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3961 ataatacgac aaggtgagga actaaaccat gaccgagtac aagcccacgg tgcgcctcgc
     4021 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4081 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4141 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4201 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4261 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4321 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4381 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4441 cgagcgcgcc ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta
     4501 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4561 gtgcatgacc cgcaagcccg gtgcctgaca cgtgctacga gatttcgatt ccaccgccgc
     4621 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4681 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4741 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4801 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4861 ctctagctag agcttggcgt aatcatggtc attaccaatg cttaatcagt gaggcaccta
     4921 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4981 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     5041 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5101 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5161 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     5221 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5281 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5341 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5401 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5461 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5521 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5581 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5641 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5701 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     5761 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5821 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     5881 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg ctcatgacca
     5941 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
     6001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6181 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6781 ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc tgacggatgg
     6841 cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt cttaagctcg
     6901 ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg taaaaacccg
     6961 cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag aatatttaag
     7021 ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg agaaaaaagc
     7081 aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata attaaactat
     7141 tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa agagaattaa
     7201 gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa attatacatg
     7261 tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt attatcattt
     7321 agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc ttcatgcaca
     7381 gtggcgttga ctcaatattg gccattagcc atattattca ttggttatat agcataaatc
     7441 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     7501 ggctcatgtc caatatgacc gccatgtt
//
