LOCUS       sequence-387048-        7708 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 7708)
  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 7708)
  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-SIGLEC11-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7708
                     /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             3284..3328
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3329..3346
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3486..3503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3492..3603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3721..3741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3726..4055
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3906..4041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3968..3987
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4103..4150
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4169..4768
                     /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(4169..4188)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4665..4685
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4898..5019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4935..4954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4989..5008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5072..5863,5864..5932))
                     /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     5695..5714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5933..6024)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6066..6085
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6091..6110
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6196..6784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6685..6704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6938..6955
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7053..7070
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7054..7070
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7526..7542
                     /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 ccttgaacaa ggaccctagc tatagcctgc aggtccagag
     1021 acaggtgcca gtgcctgaag gcctgtgtgt gatcgtgtcc tgcaacctga gctaccccag
     1081 agatggctgg gatgagtcta cagccgccta cggctactgg ttcaagggca gaacaagccc
     1141 taaaacaggc gcccctgtgg ccaccaacaa ccagagcaga gaagtggaaa tgagcacccg
     1201 ggacagattc cagctgacag gcgatcctgg caagggcagc tgtagcctgg ttatcagaga
     1261 tgcccagaga gaggacgagg cctggtactt cttcagagtg gaacggggaa gcagagtgcg
     1321 gcacagcttt ctgagcaacg cattcttcct gaaagtgacg gccctgacca agaaacccga
     1381 cgtgtacatc cccgagacac tggaacctgg acagcctgtg accgtgatct gcgtgttcaa
     1441 ctgggccttc aagaagtgcc ccgctcctag cttttcttgg actggcgctg ccctgtctcc
     1501 tcggagaaca agacctagca ccagccactt cagcgtgctg agctttaccc catctcctca
     1561 ggaccacgac accgatctga cctgccacgt ggactttagc agaaagggcg tgtcagccca
     1621 gagaaccgtc agactgagag tggcctacgc tcccaaggac ctgatcatca gcatcagcca
     1681 cgacaacaca agcgccctgg aactgcaggg caacgtgatc tacctggaag tgcagaaggg
     1741 ccagttcctg agactgctgt gtgccgctga ttctcagcct cctgctacac tgtcttgggt
     1801 gctgcaggat agagtgctga gcagctctca cccttggggc cctagaacac tgggacttga
     1861 gctgagaggc gtcagagccg gcgatagcgg cagatacaca tgcagagccg agaacagact
     1921 gggctctcag cagcaggctc tggatctgtc tgtgcagtac cctcctgaga acctgagagt
     1981 gatggtgtcc caggccaacc ggaccgtgct ggaaaatctc ggcaatggca caagcctgcc
     2041 tgtgctggaa ggacagtctc tgagactcgt gtgcgtgacc cattctagcc ctccagctag
     2101 actgagctgg accagatggg gacagacagt gggacctagc caaccttctg atcctggcgt
     2161 tctcgagctg cctcctatcc agatggaaca cgagggcgag ttcacctgtc atgcccagca
     2221 tccactcggc agccagcatg tctctctgag cctgtccgtg cactatcctc cacagctgct
     2281 gggcccatct tgttcttggg aagccgaggg cctgcactgt agctgtagtt ctcaagcctc
     2341 tcctgctcct agcctgagat ggtggctggg cgaagaactg ctcgagggca atagcagcca
     2401 gggcagcttt gaagtgacac ctagctctgc cggaccttgg gccaatagct ctctgtctct
     2461 gcatggcgga ctgtctagcg gactgagact gagatgcaag gcctggaatg tgcacggcgc
     2521 tcagtctggc tctgtgtttc agctgctgcc tggcaaactg gaacatggcg gaggactcgg
     2581 acttggcgcg ccgggcggcg acaagaccca cacctgcccc ccctgcccag ctccagaact
     2641 gctgggaggg ccttccgtgt tcctgttccc ccccaagccc aaggacaccc tgatgatcag
     2701 caggaccccc gaggtgacct gcgtggtggt ggccgtgtcc cacgaggacc cagaggtgaa
     2761 gttcaactgg tacgtggacg gcgtggaggt gcacaacgcc aagaccaagc ccagagagga
     2821 gcagtacaac agcacctaca gggtggtgtc cgtgctgacc gtgctgcacc aggactggct
     2881 gaacggcaaa gaatacaagt gcaaagtctc caacaaggcc ctggctgccc caatcgaaaa
     2941 gacaatcagc aaggccaagg gccagccacg ggagccccag gtgtacaccc tgccccccag
     3001 ccgggaggag atgaccaaga accaggtgtc cctgacctgt ctggtgaagg gcttctaccc
     3061 cagcgatatc gccgtggagt gggagagcaa cggccagccc gagaacaact acaagaccac
     3121 ccccccagtg ctggacagcg acggcagctt cttcctgtac agcaagctga ccgtggacaa
     3181 gtccaggtgg cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa
     3241 ccactacacc cagaagtccc tgagcctgag ccccggcggc agcggtctta atgatatttt
     3301 cgaggctcag aagattgagt ggcatgaaca ccaccaccat caccactaag gccggcctgt
     3361 agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc gtcatactgc
     3421 agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata gatagagaaa
     3481 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3541 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3601 atcacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3661 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3721 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3841 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3901 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3961 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4021 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4081 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     4141 ataatacgac aaggtgagga actaaaccat gaccgagtac aagcccacgg tgcgcctcgc
     4201 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4261 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4321 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4381 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4441 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4501 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4561 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4621 cgagcgcgcc ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta
     4681 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4741 gtgcatgacc cgcaagcccg gtgcctgaca cgtgctacga gatttcgatt ccaccgccgc
     4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4861 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5041 ctctagctag agcttggcgt aatcatggtc attaccaatg cttaatcagt gaggcaccta
     5101 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5161 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     5221 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5281 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5341 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     5401 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5461 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5521 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5581 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5641 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5701 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5761 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5821 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5881 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     5941 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6001 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     6061 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg ctcatgacca
     6121 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
     6181 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6241 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6301 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6361 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6421 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6661 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6721 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6781 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6841 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6901 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6961 ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc tgacggatgg
     7021 cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt cttaagctcg
     7081 ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg taaaaacccg
     7141 cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag aatatttaag
     7201 ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg agaaaaaagc
     7261 aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata attaaactat
     7321 tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa agagaattaa
     7381 gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa attatacatg
     7441 tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt attatcattt
     7501 agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc ttcatgcaca
     7561 gtggcgttga ctcaatattg gccattagcc atattattca ttggttatat agcataaatc
     7621 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     7681 ggctcatgtc caatatgacc gccatgtt
//
