LOCUS       sequence-396945-        7135 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 7135)
  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 7135)
  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-SIRPA-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7135
                     /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             2711..2755
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2756..2773
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2913..2930)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2919..3030
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3148..3168)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3153..3482
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3333..3468
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3395..3414
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3530..3577
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3596..4195
                     /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(3596..3615)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4092..4112
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4325..4446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4362..4381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4416..4435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4499..5290,5291..5359))
                     /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     5122..5141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5360..5451)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5493..5512
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5518..5537
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5623..6211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6112..6131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6365..6382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6480..6497
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6481..6497
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6953..6969
                     /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 ccgaagagga actgcaagtg atccagcctg acaagtctgt
     1021 gctggtggct gccggcgaaa cagccacact gagatgtacc gccacctctc tgatccctgt
     1081 gggccctatt cagtggttta gaggcgccgg acctggcaga gagctgatct acaatcagaa
     1141 agagggacac ttccccagag tgaccaccgt gtccgacctg accaagcgga acaacatgga
     1201 cttcagcatc cggatcggca acatcacacc agccgatgcc ggcacctact actgcgtgaa
     1261 gtttagaaag ggcagccccg acgacgtgga attcaaatct ggcgccggaa cagagctgtc
     1321 tgtgcgggct aaacctagcg ctcctgtggt ttctggacct gccgctagag ctacacctca
     1381 gcacaccgtg tcttttacct gcgagagcca cggcttcagc cccagagata tcaccctgaa
     1441 gtggttcaag aacggcaacg agctgagcga cttccagacc aatgtggacc ctgtgggaga
     1501 gagcgtgtcc tactctatcc acagcacagc caaggtggtg ctgaccagag aagatgtgca
     1561 ctcccaagtg atctgcgagg tggcccatgt gacactgcaa ggcgatcctc tgagaggcac
     1621 cgccaacctg agcgagacaa tcagagtgcc tcctacactg gaagtgaccc agcagcctgt
     1681 cagagccgag aatcaagtga acgtgacctg ccaagtgcgg aagttctacc ctcagagact
     1741 gcagctgacc tggctggaaa acggcaatgt gtccagaacc gagacagcca gcaccgtgac
     1801 agagaacaag gacggcacat acaactggat gagctggctg ctggtcaatg tgtctgccca
     1861 cagagatgac gtgaagctga catgccaggt ggaacacgat ggacagcctg ccgtgtctaa
     1921 gagccacgac ctgaaagtgt ctgctcaccc caaagagcag ggctctaaca cagccgccga
     1981 gaacacaggc agcaacgagc ggaacatcta tggcgcgccg ggcggcgaca agacccacac
     2041 ctgccccccc tgcccagctc cagaactgct gggagggcct tccgtgttcc tgttcccccc
     2101 caagcccaag gacaccctga tgatcagcag gacccccgag gtgacctgcg tggtggtggc
     2161 cgtgtcccac gaggacccag aggtgaagtt caactggtac gtggacggcg tggaggtgca
     2221 caacgccaag accaagccca gagaggagca gtacaacagc acctacaggg tggtgtccgt
     2281 gctgaccgtg ctgcaccagg actggctgaa cggcaaagaa tacaagtgca aagtctccaa
     2341 caaggccctg gctgccccaa tcgaaaagac aatcagcaag gccaagggcc agccacggga
     2401 gccccaggtg tacaccctgc cccccagccg ggaggagatg accaagaacc aggtgtccct
     2461 gacctgtctg gtgaagggct tctaccccag cgatatcgcc gtggagtggg agagcaacgg
     2521 ccagcccgag aacaactaca agaccacccc cccagtgctg gacagcgacg gcagcttctt
     2581 cctgtacagc aagctgaccg tggacaagtc caggtggcag cagggcaacg tgttcagctg
     2641 cagcgtgatg cacgaggccc tgcacaacca ctacacccag aagtccctga gcctgagccc
     2701 cggcggcagc ggtcttaatg atattttcga ggctcagaag attgagtggc atgaacacca
     2761 ccaccatcac cactaaggcc ggcctgtagt cactggcaca acgcgttacc cggtaagcca
     2821 atcgggtata cacggtcgtc atactgcaga cagggttctt ctactttgca agatagtctt
     2881 gagtagtaaa ataaatagat agagaaaatc agcctcgact gtgccttcta gttgccagcc
     2941 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3001 cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct
     3061 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3121 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3361 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3421 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3481 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     3541 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac
     3601 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3661 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3721 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3781 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3841 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3901 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3961 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4021 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4081 atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4141 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt
     4201 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4321 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4441 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt
     4501 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4561 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4621 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
     4681 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4741 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4801 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4861 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4921 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4981 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5041 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5101 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5161 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5221 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5281 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5341 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5401 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
     5461 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     5521 gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5761 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6361 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     6421 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     6481 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     6541 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     6601 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     6661 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     6721 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     6781 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     6841 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     6901 agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt aattgtgagc
     6961 ggataacaat tacgagcttc atgcacagtg gcgttgactc aatattggcc attagccata
     7021 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7081 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
