LOCUS       sequence-368849-        7933 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 7933)
  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 7933)
  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-CD300C-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(140..187)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(235..564)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(303..322)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     549..569
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    687..798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     787..804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(944..961)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(962..1006)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2312..2331)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2344..2362)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2345..2362)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2572..2596)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2593..2796)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2622..2642)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2797..3176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    4681..4697
                     /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)."
     primer_bind     complement(5153..5170)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5153..5169)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5439..6027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5519..5538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6113..6132)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(6138..6157)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        6199..6290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6291..6359,6360..7151)
                     /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     complement(6509..6528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7204..7325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7215..7234)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7269..7288
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(7455..7933,1..121))
                     /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(7538..7558)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
       61 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
      121 tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat
      181 tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
      241 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc
      301 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
      361 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      421 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      481 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      541 ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa
      601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      661 tagaccgaga tagggttgag tgttgtgatg caatttcctc attttattag gaaaggacag
      721 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      781 ggcaactaga aggcacagtc gaggctgatt ttctctatct atttatttta ctactcaaga
      841 ctatcttgca aagtagaaga accctgtctg cagtatgacg accgtgtata cccgattggc
      901 ttaccgggta acgcgttgtg ccagtgacta caggccggcc ttagtggtga tggtggtggt
      961 gttcatgcca ctcaatcttc tgagcctcga aaatatcatt aagaccgctg ccgccggggc
     1021 tcaggctcag ggacttctgg gtgtagtggt tgtgcagggc ctcgtgcatc acgctgcagc
     1081 tgaacacgtt gccctgctgc cacctggact tgtccacggt cagcttgctg tacaggaaga
     1141 agctgccgtc gctgtccagc actggggggg tggtcttgta gttgttctcg ggctggccgt
     1201 tgctctccca ctccacggcg atatcgctgg ggtagaagcc cttcaccaga caggtcaggg
     1261 acacctggtt cttggtcatc tcctcccggc tggggggcag ggtgtacacc tggggctccc
     1321 gtggctggcc cttggccttg ctgattgtct tttcgattgg ggcagccagg gccttgttgg
     1381 agactttgca cttgtattct ttgccgttca gccagtcctg gtgcagcacg gtcagcacgg
     1441 acaccaccct gtaggtgctg ttgtactgct cctctctggg cttggtcttg gcgttgtgca
     1501 cctccacgcc gtccacgtac cagttgaact tcacctctgg gtcctcgtgg gacacggcca
     1561 ccaccacgca ggtcacctcg ggggtcctgc tgatcatcag ggtgtccttg ggcttggggg
     1621 ggaacaggaa cacggaaggc cctcccagca gttctggagc tgggcagggg gggcaggtgt
     1681 gggtcttgtc gccgcccggc gcgcctcgga cgttgctgaa caggctgcca ggatgtggag
     1741 aaggttcggg gctatccttt ctggtcacgc tgggccatgt gtgcacaggc agctttgtag
     1801 gtgggccaga tgtgcccatg ctagactgtg gagagctggc tgttgttgtg ccggctggaa
     1861 acacggacac ttccacttcc acgatggggt cgtgaaagtc tctcagccaa ggggtgtcca
     1921 ctccgcacca atatgtgccg gcatcttcct cggtcaggtt ttccagtgtc acggtgaagc
     1981 tcaggttggc ggggctgtct ctgatagaca ctctgccgtt tctcttgccg gcgctgccct
     2041 ttgtttccac gatcttgtcg catctcagga tctgaggagg ccggcaccag aacttgttca
     2101 gggttctgtg ctctttctcg tatctgcact gcacgctcag agatccgcca acaggtccgg
     2161 ccactgtcat agggtgagac aggggaaaat atccggcggc cgcagcgaac accaggcaga
     2221 agatgtagct cagggcaatg atggtcttca tggtggcgct agcaggcgcg ttttcgatag
     2281 aaccgaggtg cagttggacc tgggagtgga cacctgtgga gagaaaggca aagtggatgt
     2341 tattctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta
     2401 gccgcgttct aacgaagcca agggggtggg cctatagact ctataggcgg tacttacgtc
     2461 actcttggca cggggaatcc gcgttccaat gcaccgttcc cggccgcgga ggctggatcg
     2521 gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg
     2581 gttcactaaa cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     2641 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     2701 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     2761 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     2821 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     2881 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     2941 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     3001 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     3061 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     3121 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgccaaca
     3181 tggcggtcat attggacatg agccaatata aatgtacata ttatgatata gatacaacgt
     3241 actgatgaat cccctaatga ttttggtaaa aatcattaag ttaaggtgga tacacatctt
     3301 gtcatatgat caaatggttt cgcgaaaaat caataatcag acaacaagat gtgcgaactc
     3361 gatattttac acgactctct ttaccaattc tgccccgaat tacacttaaa acgactcaac
     3421 agcttaacgt tggcttgcca cgcattactt gactgtaaaa ctctcactct taccgaactt
     3481 ggccgtaacc tgccaaccaa agcgagaaca aaacataaca tcaaacgaat cgaccgattg
     3541 ttaggtaatc gtcacctcca caaagagcga ctcgctgtat accgttggca tgctagcttt
     3601 atctgttcgg gcaatacgat gcccattgta cttgttgact ggtctgatat tcgtgagcaa
     3661 aaacgactta tggtattgcg agcttcagtc gcactacacg gtcgttctgt tactctttat
     3721 gagaaagcgt tcccgctttc agagcaatgt tcaaagaaag ctcatgacca atttctagcc
     3781 gaccttgcga gcattctacc gagtaacacc acaccgctca ttgtcagtga tgctggcttt
     3841 aaagtgccat ggtataaatc cgttgagaag ctgggttggt actggttaag tcgagtaaga
     3901 ggaaaagtac aatatgcaga cctaggagcg gaaaactgga aacctatcag caacttacat
     3961 gatatgtcat ctagtcactc aaagacttta ggctataaga ggctgactaa aagcaatcca
     4021 atctcatgcc aaattctatt gtataaatct cgctctaaag gccgaaaaaa tcagcgctcg
     4081 acacggactc attgtcacca cccgtcacct aaaatctact cagcgtcggc aaaggagcca
     4141 tgggttctag caactaactt acctgttgaa attcgaacac ccaaacaact tgttaatatc
     4201 tattcgaagc gaatgcagat tgaagaaacc ttccgagact tgaaaagtcc tgcctacgga
     4261 ctaggcctac gccatagccg aacgagcagc tcagagcgtt ttgatatcat gctgctaatc
     4321 gccctgatgc ttcaactaac atgttggctt gcgggcgttc atgctcagaa acaaggttgg
     4381 gacaagcact tccaggctaa cacagtcaga aatcgaaacg tactctcaac agttcgctta
     4441 ggcatggaag ttttgcggca ttctggctac acaataacaa gggaagactt actcgtggct
     4501 gcaaccctac tagctcaaaa tttattcaca catggttacg ctttggggaa attatgaggg
     4561 gatctctcag tacaacgtat gcaatggcca atagccaata ttgatttatg ctatataacc
     4621 aatgaataat atggctaatg gccaatattg agtcaacgcc actgtgcatg aagctcgtaa
     4681 ttgttatccg ctcacaatta agggcgacac aaaatttatt ctaaatgata ataaatactg
     4741 ataacatctt atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat
     4801 caaaaactga ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa
     4861 ctagaaatat tgtatataca aaaaatcata aataatagat gaatagttta attataggtg
     4921 ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata
     4981 aggttaaata attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa
     5041 tgctctttcc ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt
     5101 gcggattaac gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg
     5161 tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt
     5221 ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac
     5281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5821 tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga
     5881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6061 gctcagtgga acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcaaggggt
     6121 gttatgagcc atattcaggt ataaatgggc tcgcgataat gttcagaatt ggttaattgg
     6181 ttgtaacact gacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6241 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat atgagtattc
     6301 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6361 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6421 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6481 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6541 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6601 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6661 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6721 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6781 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcga
     6841 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6901 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6961 cggctggctg gtttattgct gataaatccg gagccggtga gcgtggttct cgcggtatca
     7021 tcgcagcgct ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7081 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7141 agcattggta atgaccatga ttacgccaag ctctagctag aggtcgacgg tatacagaca
     7201 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     7261 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     7321 aagttggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     7381 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     7441 tctcgtagca cgtgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     7501 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     7561 ggcgcggatg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     7621 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     7681 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     7741 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     7801 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     7861 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     7921 cgctcgatgt ggc
//
