LOCUS       sequence-386096-        7465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7465)
  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 7465)
  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-ICAM1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7465
                     /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             3041..3085
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3086..3103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3243..3260)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3249..3360
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3478..3498)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3483..3812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3663..3798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3725..3744
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3860..3907
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3926..4525
                     /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(3926..3945)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4422..4442
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4655..4776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4692..4711)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4746..4765
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4829..5620,5621..5689))
                     /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     5452..5471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5690..5781)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5823..5842
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5848..5867
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5953..6541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6442..6461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6695..6712
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6810..6827
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6811..6827
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7283..7299
                     /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 cccaaacatc tgtgtcccca agcaaagtga tcctgcctag
     1021 aggcggcagc gtgctggtca catgtagcac ctcttgcgac cagcctaagc tgctgggcat
     1081 cgagacaccc ctgcctaaga aagagctgct gctgcccggc aacaaccgga aggtgtacga
     1141 gctgagcaac gtgcaagagg acagccagcc tatgtgctac agcaactgcc ctgatggcca
     1201 gtctaccgcc aagaccttcc tgaccgtgta ctggacccct gagagagtgg aactggcccc
     1261 tctgccttct tggcagcctg tgggcaagaa tctgaccctg agatgccagg tggaaggcgg
     1321 agcccctaga gccaatctga cagtggttct gctgcggggc gagaaagaac tgaagaggga
     1381 acctgccgtg ggcgagcctg ctgaagtgac aacaacagtg ctcgtgcgga gagatcacca
     1441 cggcgccaat ttcagctgcc ggacagaact ggatctgagg ccacagggcc tcgagctgtt
     1501 cgagaatacc agcgctcctt accagctgca gaccttcgtg ctgcctgcta cacctcctca
     1561 gctggtgtct cccagagtgc tggaagtgga tacccagggc acagtcgtgt gcagcctgga
     1621 tggcctgttt cctgtgtctg aggcccaagt gcatctggcc ctgggagatc agagactgaa
     1681 ccccacagtg acctacggca acgacagctt ttctgccaag gccagcgtgt ccgtgacagc
     1741 cgaagatgag ggaacccaga ggctgacctg tgccgtgatc ctgggcaacc agagccaaga
     1801 gacactgcag accgtgacca tctacagctt ccccgctcct aatgtgatcc tgaccaagcc
     1861 tgaggtgtcc gagggcaccg aagtgacagt gaagtgcgag gcccatcctc gggccaaagt
     1921 gacactgaat ggcgtgccag ctcagccact gggacctaga gcacagctcc tgctgaaagc
     1981 cacacctgag gacaacggca gaagcttcag ctgcagcgcc acactggaag tcgccggaca
     2041 gctgatccac aagaaccaga ccagagaact gagagtgctg tacggcccca gactggacga
     2101 gagagattgc cccggaaatt ggacctggcc tgagaacagc cagcagaccc ctatgtgtca
     2161 ggcctgggga aatcctctgc ctgagctgaa gtgcctgaag gacggcacct ttcctctgcc
     2221 aatcggcgag tctgtgaccg tgacaagaga tctggaaggc acctacctgt gcagagccag
     2281 atccacacag ggcgaagtga cccggaaagt gaccgtcaat gtgctgagcc ccagatacga
     2341 aggcgcgccg ggcggcgaca agacccacac ctgccccccc tgcccagctc cagaactgct
     2401 gggagggcct tccgtgttcc tgttcccccc caagcccaag gacaccctga tgatcagcag
     2461 gacccccgag gtgacctgcg tggtggtggc cgtgtcccac gaggacccag aggtgaagtt
     2521 caactggtac gtggacggcg tggaggtgca caacgccaag accaagccca gagaggagca
     2581 gtacaacagc acctacaggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa
     2641 cggcaaagaa tacaagtgca aagtctccaa caaggccctg gctgccccaa tcgaaaagac
     2701 aatcagcaag gccaagggcc agccacggga gccccaggtg tacaccctgc cccccagccg
     2761 ggaggagatg accaagaacc aggtgtccct gacctgtctg gtgaagggct tctaccccag
     2821 cgatatcgcc gtggagtggg agagcaacgg ccagcccgag aacaactaca agaccacccc
     2881 cccagtgctg gacagcgacg gcagcttctt cctgtacagc aagctgaccg tggacaagtc
     2941 caggtggcag cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca
     3001 ctacacccag aagtccctga gcctgagccc cggcggcagc ggtcttaatg atattttcga
     3061 ggctcagaag attgagtggc atgaacacca ccaccatcac cactaaggcc ggcctgtagt
     3121 cactggcaca acgcgttacc cggtaagcca atcgggtata cacggtcgtc atactgcaga
     3181 cagggttctt ctactttgca agatagtctt gagtagtaaa ataaatagat agagaaaatc
     3241 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3301 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3361 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3421 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3481 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3661 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3721 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3781 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3841 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3901 atacgacaag gtgaggaact aaaccatgac cgagtacaag cccacggtgc gcctcgccac
     3961 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4021 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4081 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4141 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4201 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4261 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4321 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4381 gcgcgccggg gtgcccgcct tcctggagac atccgcgccc cgcaacctcc ccttctacga
     4441 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4501 catgacccgc aagcccggtg cctgacacgt gctacgagat ttcgattcca ccgccgcctt
     4561 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4621 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4681 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4741 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4801 tagctagagc ttggcgtaat catggtcatt accaatgctt aatcagtgag gcacctatct
     4861 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4921 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     4981 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5041 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5101 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     5161 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5221 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5281 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5341 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5401 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5461 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5521 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5581 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5641 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt
     5701 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5761 gtatttagaa aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca
     5821 ttatcgcgag cccatttata cctgaatatg gctcataaca ccccttgctc atgaccaaaa
     5881 tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     5941 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6001 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6061 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6121 agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6541 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6601 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6661 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6721 aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga cggatggcct
     6781 ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc
     6841 cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa aaacccgctt
     6901 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat atttaagggc
     6961 gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga aaaaagcaac
     7021 gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt aaactattca
     7081 tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa
     7141 aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca
     7201 acgataatac aaaatataat acaaactata agatgttatc agtatttatt atcatttaga
     7261 ataaattttg tgtcgccctt aattgtgagc ggataacaat tacgagcttc atgcacagtg
     7321 gcgttgactc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     7381 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     7441 tcatgtccaa tatgaccgcc atgtt
//
