LOCUS       sequence-368391-        6547 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 6547)
  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 6547)
  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-CD300LD-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(65..156)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     198..217
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     223..242
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      328..916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     817..836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1070..1087
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1185..1202
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1186..1202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1658..1674
                     /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        1841..2220
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2221..2424
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2375..2395
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2421..2445
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2655..2673
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2655..2672
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2686..2705
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3963..4007
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4008..4025
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4165..4182)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4171..4282
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4400..4420)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4405..4734
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4585..4720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4647..4666
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4782..4829
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4848..5447
                     /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(4848..4867)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5344..5364
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5577..5698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5614..5633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5668..5687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5751..6542,6543..6547,1..64))
                     /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     6374..6393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
       61 tcatattctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      121 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt acaaccaatt
      181 aaccaattct gaacattatc gcgagcccat ttatacctga atatggctca taacacccct
      241 tgctcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
      301 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      361 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      421 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
      481 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
      541 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      601 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      661 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      721 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      781 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      841 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      901 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      961 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1021 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1081 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     1141 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     1201 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     1261 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     1321 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     1381 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     1441 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     1501 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     1561 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     1621 ttattatcat ttagaataaa ttttgtgtcg cccttaattg tgagcggata acaattacga
     1681 gcttcatgca cagtggcgtt gactcaatat tggccattag ccatattatt cattggttat
     1741 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     1801 tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt
     1861 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1921 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1981 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2041 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2101 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2161 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2221 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2281 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2341 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2401 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2461 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
     2521 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
     2581 tataggccca cccccttggc ttcgttagaa cgcggctaca attaatacat aaccttatgt
     2641 atcatacaca tacgatttag gtgacactat agaataacat ccactttgcc tttctctcca
     2701 caggtgtcca ctcccaggtc caactgcacc tcggttctat cgaaaacgcg cctgctagcg
     2761 ccaccatgaa gaccatcatt gccctgagct acatcttctg cctggtgttc gctgcggccg
     2821 ccgcaaagat cacaggccct acaacagtga atggcagcga gcagggaagc ctgacagtgc
     2881 agtgtgccta tggcagcggc tgggagacat atctgaaatg gcgttgtcag ggcgccgact
     2941 ggaactactg caacatcctg gtcaagacca acggctccga gcaagaagtg aagaaaaacc
     3001 gggtgtccat ccgggacaac cagaaaaacc acgtgttcac cgtgaccatg gaaaacctga
     3061 agagggacga cgccgacagc tattggtgcg gcacagaaag acctggcatc gacctgggag
     3121 tgaaggtgca agtgaccatc aatcctggca cacagaccgc cgtgtctgag tggacaacaa
     3181 caacagccag cctggccttt accgctgccg ccacacagaa aacaagcagc cctctgacca
     3241 gatctcccct gaagtctacc catggcgcgc cgggcggcga caagacccac acctgccccc
     3301 cctgcccagc tccagaactg ctgggagggc cttccgtgtt cctgttcccc cccaagccca
     3361 aggacaccct gatgatcagc aggacccccg aggtgacctg cgtggtggtg gccgtgtccc
     3421 acgaggaccc agaggtgaag ttcaactggt acgtggacgg cgtggaggtg cacaacgcca
     3481 agaccaagcc cagagaggag cagtacaaca gcacctacag ggtggtgtcc gtgctgaccg
     3541 tgctgcacca ggactggctg aacggcaaag aatacaagtg caaagtctcc aacaaggccc
     3601 tggctgcccc aatcgaaaag acaatcagca aggccaaggg ccagccacgg gagccccagg
     3661 tgtacaccct gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc ctgacctgtc
     3721 tggtgaaggg cttctacccc agcgatatcg ccgtggagtg ggagagcaac ggccagcccg
     3781 agaacaacta caagaccacc cccccagtgc tggacagcga cggcagcttc ttcctgtaca
     3841 gcaagctgac cgtggacaag tccaggtggc agcagggcaa cgtgttcagc tgcagcgtga
     3901 tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgagc cccggcggca
     3961 gcggtcttaa tgatattttc gaggctcaga agattgagtg gcatgaacac caccaccatc
     4021 accactaagg ccggcctgta gtcactggca caacgcgtta cccggtaagc caatcgggta
     4081 tacacggtcg tcatactgca gacagggttc ttctactttg caagatagtc ttgagtagta
     4141 aaataaatag atagagaaaa tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4201 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4261 aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt cttttgattt
     4321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4501 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4561 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4621 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4681 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4741 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     4801 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg accgagtaca
     4861 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     4921 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4981 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5041 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     5101 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     5161 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     5221 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     5281 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acatccgcgc
     5341 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     5401 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac gtgctacgag
     5461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5701 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca ttaccaatgc
     5761 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5821 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagcgctgcg
     5881 atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa ccagccagcc
     5941 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6001 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6061 atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6361 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6541 tgagcaa
//
