LOCUS       sequence-492991-        8140 bp    DNA     circular SYN 18-NOV-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 8140)
  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 8140)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 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-EDA-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..111)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        96..425
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      276..411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     338..357
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        473..520
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             539..1138
                     /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(539..558)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1035..1055
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1268..1389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1305..1324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1359..1378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1442..2233,2234..2302))
                     /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     2065..2084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2303..2394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2436..2455
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2461..2480
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2566..3154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3055..3074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3308..3325
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3423..3440
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3424..3440
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3896..3912
                     /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)."
     enhancer        4079..4458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4459..4662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4613..4633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4659..4683
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4893..4911
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4893..4910
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4924..4943
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6102..6125
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             7785..7829
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             7830..7856
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(7996..8013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8002..8113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
       61 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      361 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      421 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
      481 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
      541 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
      601 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
      661 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
      721 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
      781 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
      841 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
      901 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
      961 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     1021 gacatccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     1081 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaca
     1141 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     1201 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     1261 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1321 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1381 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     1441 attaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1501 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1561 ccagcgctgc gatgataccg cgagaaccac gctcaccggc tccggattta tcagcaataa
     1621 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1681 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1741 acgttgttgc catcgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1801 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1861 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1921 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1981 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2041 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2101 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2161 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2221 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2281 cacggaaatg ttgaatactc atattcttcc tttttcaata ttattgaagc atttatcagg
     2341 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2401 tcagtgttac aaccaattaa ccaattctga acattatcgc gagcccattt atacctgaat
     2461 atggctcata acaccccttg ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     2521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2701 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
     2761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3181 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3241 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3301 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
     3361 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     3421 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     3481 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     3541 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     3601 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     3661 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     3721 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     3781 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     3841 ataagatgtt atcagtattt attatcattt agaataaatt ttgtgtcgcc cttaattgtg
     3901 agcggataac aattacgagc ttcatgcaca gtggcgttga ctcaatattg gccattagcc
     3961 atattattca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     4021 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg
     4081 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4141 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4201 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4261 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4321 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4381 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4441 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     4501 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     4561 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     4621 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     4681 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     4741 agcctccgcg gccgggaacg gtgcattgga acgcggattc cccgtgccaa gagtgacgta
     4801 agtaccgcct atagagtcta taggcccacc cccttggctt cgttagaacg cggctacaat
     4861 taatacataa ccttatgtat catacacata cgatttaggt gacactatag aataacatcc
     4921 actttgcctt tctctccaca ggtgtccact cccaggtcca actgcacctc ggttctatcg
     4981 aaaacgcgcc tgctagcgcc accatgaaga ccatcattgc cctgagctac atcttctgcc
     5041 tggtgttcgc tgcggccgcc gaactgagaa gcgagcttag aagagagaga ggcgccgaga
     5101 gcagactcgg aggatctgga actcctggca caagcggaac actgtctagc cttggaggac
     5161 tggaccctga cagccctatc acatctcacc tgggccagcc ttctccaaag cagcaacctc
     5221 ttgaacctgg cgaggccgct ctgcacagtg attctcagga tggacatcag atggccctgc
     5281 tgaacttctt cttccccgac gaaaaaccct acagcgaaga ggaatccaga agagtgcggc
     5341 ggaacaagcg gagcaagagc aatgaaggtg ccgacggacc cgtgaagaac aagaagaagg
     5401 gcaagaaggc cggacctcct ggacctaatg gaccaccagg tcctccaggt ccacctggac
     5461 cacaaggacc acctggaata cctggcatcc ctggaatccc aggcacaaca gttatgggac
     5521 ctccagggcc gcctgggcct ccaggacctc aaggccctcc tggcttgcaa ggaccttctg
     5581 gtgctgccga taaggccggc accagagaaa atcagcctgc cgtggttcat ctgcaaggcc
     5641 agggatctgc catccaagtg aagaacgatc tgtctggcgg cgtgctgaac gactggtcca
     5701 gaatcaccat gaatcccaag gtgttcaagc tgcaccccag atccggcgaa ctggaagtgc
     5761 tggtggatgg cacctacttc atctacagcc aggtggaagt gtactacatc aacttcaccg
     5821 acttcgccag ctacgaggtg gtggtggatg agaagccttt cctgcagtgc accagatcca
     5881 tcgagacagg caagaccaac tacaatacct gctacacagc cggcgtgtgc ctgctgaagg
     5941 caagacagaa aatcgccgtg aagatggtgc acgccgacat cagcatcaac atgagcaagc
     6001 acaccacctt tttcggcgcc atcagactgg gagaagcccc tgcttctggc gcgccggaag
     6061 atcaagttga tccaagattg attgatggca agggcggatc tctggaggtg ctgttccagg
     6121 gaccaggcgg aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc
     6181 aggagaccaa tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga
     6241 tcaccttcct gaagaacacc gtgatggagt gcgacgcctg cggtatgcag ccctctagag
     6301 gctccggctc catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag
     6361 ccgaggccct gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca
     6421 tcatcttcct gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag
     6481 ggcagaagaa ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg
     6541 tggctctgtc caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca
     6601 cggcctacct gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc
     6661 gctttaacca gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca
     6721 agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag
     6781 ccggcaccta cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct
     6841 cggcccgcca ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg
     6901 acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt
     6961 acccagatga ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat
     7021 ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt
     7081 ccctggaccc gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg
     7141 agatccaccg agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc
     7201 gcctgctgag caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc
     7261 atggtcatca tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg
     7321 ccattgagag ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg
     7381 accactccca cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc
     7441 tggcccctgg caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc
     7501 caggctatgt gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc
     7561 ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg
     7621 tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct
     7681 tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg
     7741 cgccccccgc cggcaccacc gacgccgcgc acccgggtgg cggaggactg aacgacatct
     7801 tcgaggctca gaaaatcgaa tggcacgaac atcaccatca ccatcaccat caccattaag
     7861 gccggcctgt agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc
     7921 gtcatactgc agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata
     7981 gatagagaaa atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     8041 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     8101 aggaaattgc atcacaacac tcaaccctat ctcggtctat
//
