LOCUS       sequence-368828-        6469 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 6469)
  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 6469)
  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-NFAM1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     215..235
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    353..464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     453..470
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(610..627)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(628..672)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1852..1871)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1884..1902)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1885..1902)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2112..2136)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2133..2336)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2162..2182)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2337..2716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2883..2899
                     /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(3355..3372)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3355..3371)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3470..3487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3641..4229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3721..3740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4315..4334)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4340..4359)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4401..4492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4493..4561,4562..5353)
                     /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(4711..4730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5406..5527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5417..5436)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5471..5490
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5657..6256)
                     /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(5740..5760)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6237..6256
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6275..6322)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6370..6469,1..230))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6384..6469,1..50)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6438..6457)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
       61 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      121 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      181 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
      241 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      301 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gtgatgcaat
      361 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      421 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgattttct
      481 ctatctattt attttactac tcaagactat cttgcaaagt agaagaaccc tgtctgcagt
      541 atgacgaccg tgtatacccg attggcttac cgggtaacgc gttgtgccag tgactacagg
      601 ccggccttag tggtgatggt ggtggtgttc atgccactca atcttctgag cctcgaaaat
      661 atcattaaga ccgctgccgc cggggctcag gctcagggac ttctgggtgt agtggttgtg
      721 cagggcctcg tgcatcacgc tgcagctgaa cacgttgccc tgctgccacc tggacttgtc
      781 cacggtcagc ttgctgtaca ggaagaagct gccgtcgctg tccagcactg ggggggtggt
      841 cttgtagttg ttctcgggct ggccgttgct ctcccactcc acggcgatat cgctggggta
      901 gaagcccttc accagacagg tcagggacac ctggttcttg gtcatctcct cccggctggg
      961 gggcagggtg tacacctggg gctcccgtgg ctggcccttg gccttgctga ttgtcttttc
     1021 gattggggca gccagggcct tgttggagac tttgcacttg tattctttgc cgttcagcca
     1081 gtcctggtgc agcacggtca gcacggacac caccctgtag gtgctgttgt actgctcctc
     1141 tctgggcttg gtcttggcgt tgtgcacctc cacgccgtcc acgtaccagt tgaacttcac
     1201 ctctgggtcc tcgtgggaca cggccaccac cacgcaggtc acctcggggg tcctgctgat
     1261 catcagggtg tccttgggct tgggggggaa caggaacacg gaaggccctc ccagcagttc
     1321 tggagctggg cagggggggc aggtgtgggt cttgtcgccg cccggcgcgc ctttttgagg
     1381 gctctgagga ggctctctgt agccggcatc tctgacgagg atgaaggttc cagagcctct
     1441 gactgtgctg tggggccagt gcacagaaca gtaatatgtg cctgtggcag aggcgccagg
     1501 aagcaccagg gtcacttggc aatccagtgt gtgagactgg ttctcggtgc ccaggccagg
     1561 gtgacaattt gtaggcttct tggggcttct ctggccctgc aggtcctcgt ggaagtagga
     1621 cacggtgaac accttgaact ggggtgtgta ggggtatgtg atccggcagc tgaagctgat
     1681 ggcggtgttg gccagagagg ccataatagg cagtcctgtg tgtgtcacag attgggcggc
     1741 cgcagcgaac accaggcaga agatgtagct cagggcaatg atggtcttca tggtggcgct
     1801 agcaggcgcg ttttcgatag aaccgaggtg cagttggacc tgggagtgga cacctgtgga
     1861 gagaaaggca aagtggatgt tattctatag tgtcacctaa atcgtatgtg tatgatacat
     1921 aaggttatgt attaattgta gccgcgttct aacgaagcca agggggtggg cctatagact
     1981 ctataggcgg tacttacgtc actcttggca cggggaatcc gcgttccaat gcaccgttcc
     2041 cggccgcgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg
     2101 cgtctccagg cgatctgacg gttcactaaa cgagctctgc ttatatagac ctcccaccgt
     2161 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     2221 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     2281 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     2341 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     2401 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     2461 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     2521 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     2581 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     2641 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     2701 gtcaataatc aatgccaaca tggcggtcat attggacatg agccaatata aatgtacata
     2761 ttatgatata gatacaacgt atgcaatggc caatagccaa tattgattta tgctatataa
     2821 ccaatgaata atatggctaa tggccaatat tgagtcaacg ccactgtgca tgaagctcgt
     2881 aattgttatc cgctcacaat taagggcgac acaaaattta ttctaaatga taataaatac
     2941 tgataacatc ttatagtttg tattatattt tgtattatcg ttgacatgta taattttgat
     3001 atcaaaaact gattttccct ttattatttt cgagatttat tttcttaatt ctctttaaca
     3061 aactagaaat attgtatata caaaaaatca taaataatag atgaatagtt taattatagg
     3121 tgttcatcaa tcgaaaaagc aacgtatctt atttaaagtg cgttgctttt ttctcattta
     3181 taaggttaaa taattctcat atatcaagca aagtgacagg cgcccttaaa tattctgaca
     3241 aatgctcttt ccctaaactc cccccataaa aaaacccgcc gaagcgggtt tttacgttat
     3301 ttgcggatta acgattactc gttatcagaa ccgcccaggg ggcccgagct taagactggc
     3361 cgtcgtttta caacacagaa agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc
     3421 ttctgcttag tttgatgcct ggcagttccc tactctcgcc ttccgcttcc tcgctcactg
     3481 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3541 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4021 ggtatgtagg cggtgctaca gagttcttga agtggtgggc taactacggc tacactagaa
     4081 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4261 acgctcagtg gaacgacgcg cgcgtaactc acgttaaggg attttggtca tgagcaaggg
     4321 gtgttatgag ccatattcag gtataaatgg gctcgcgata atgttcagaa ttggttaatt
     4381 ggttgtaaca ctgaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga atatgagtat
     4501 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4561 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4621 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4681 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4741 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4801 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4861 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4921 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4981 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5041 gatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5101 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5161 tccggctggc tggtttattg ctgataaatc cggagccggt gagcgtggtt ctcgcggtat
     5221 catcgcagcg ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5281 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5341 taagcattgg taatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     5401 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     5461 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5521 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     5581 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     5641 aatctcgtag cacgtgtcag gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt
     5701 cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgc
     5761 ggggcgcgga tgtctccagg aaggcgggca ccccggcgcg ctcggccgcc tccactccgg
     5821 ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg
     5881 ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc atctgttgct
     5941 gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg
     6001 cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga
     6061 cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga
     6121 cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg
     6181 cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg
     6241 gcttgtactc ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact
     6301 atgccgatga ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc
     6361 cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     6421 ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagt
//
