LOCUS       sequence-368832-        6481 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 6481)
  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 6481)
  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-CD83-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6481
                     /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(2198..2217)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2230..2248)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2231..2248)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2458..2482)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2479..2682)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2508..2528)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2683..3062
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3229..3245
                     /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(3701..3718)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3701..3717)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3816..3833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4661..4680)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4686..4705)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4747..4838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4839..4907,4908..5699)
                     /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(5057..5076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5752..5873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5763..5782)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5817..5836
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6003..6481,1..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(6086..6106)
                     /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 gcgccttcgg cgcggtactt cttgaaggtt tcctctttcc
     1741 gctgagcggg acagccggtc actctcagga tcactttgcc gctcagattt ctctggccat
     1801 cggggtcctg cagggtacat ctgtaggtgc cgctgttgca gctggtggtg ttccggatct
     1861 tcaggctgta gggcctctca ttaggggcgt cgaagctgcc gttctggcct ttctggtggt
     1921 agtgctggcc tctcaggtga tcttcttgag gggtttccat ccgttcctcg ccgccttcca
     1981 gcagcttaac ccaagacacg gtgtatggca cctgagggtc ccaaggagct gtacaaggca
     2041 ggtccacgtc ctcgctacag gccactttca cttcaggtgt ggcggccgca gcgaacacca
     2101 ggcagaagat gtagctcagg gcaatgatgg tcttcatggt ggcgctagca ggcgcgtttt
     2161 cgatagaacc gaggtgcagt tggacctggg agtggacacc tgtggagaga aaggcaaagt
     2221 ggatgttatt ctatagtgtc acctaaatcg tatgtgtatg atacataagg ttatgtatta
     2281 attgtagccg cgttctaacg aagccaaggg ggtgggccta tagactctat aggcggtact
     2341 tacgtcactc ttggcacggg gaatccgcgt tccaatgcac cgttcccggc cgcggaggct
     2401 ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat
     2461 ctgacggttc actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     2521 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     2581 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     2641 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     2701 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     2761 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     2821 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     2881 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     2941 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     3001 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     3061 ccaacatggc ggtcatattg gacatgagcc aatataaatg tacatattat gatatagata
     3121 caacgtatgc aatggccaat agccaatatt gatttatgct atataaccaa tgaataatat
     3181 ggctaatggc caatattgag tcaacgccac tgtgcatgaa gctcgtaatt gttatccgct
     3241 cacaattaag ggcgacacaa aatttattct aaatgataat aaatactgat aacatcttat
     3301 agtttgtatt atattttgta ttatcgttga catgtataat tttgatatca aaaactgatt
     3361 ttccctttat tattttcgag atttattttc ttaattctct ttaacaaact agaaatattg
     3421 tatatacaaa aaatcataaa taatagatga atagtttaat tataggtgtt catcaatcga
     3481 aaaagcaacg tatcttattt aaagtgcgtt gcttttttct catttataag gttaaataat
     3541 tctcatatat caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg ctctttccct
     3601 aaactccccc cataaaaaaa cccgccgaag cgggttttta cgttatttgc ggattaacga
     3661 ttactcgtta tcagaaccgc ccagggggcc cgagcttaag actggccgtc gttttacaac
     3721 acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct gcttagtttg
     3781 atgcctggca gttccctact ctcgccttcc gcttcctcgc tcactgactc gctgcgctcg
     3841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtgggctaac tacggctaca ctagaagaac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gacgcgcgcg taactcacgt taagggattt tggtcatgag caaggggtgt tatgagccat
     4681 attcaggtat aaatgggctc gcgataatgt tcagaattgg ttaattggtt gtaacactga
     4741 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4801 cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagtattcaa catttccgtg
     4861 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4921 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4981 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5041 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5101 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5161 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5221 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5281 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5341 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcgatg gcaacaacgt
     5401 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5461 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5521 ttattgctga taaatccgga gccggtgagc gtggttctcg cggtatcatc gcagcgctgg
     5581 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5641 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat
     5701 gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca
     5761 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5821 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg
     5881 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     5941 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg
     6001 tgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     6061 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggatgtc
     6121 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
     6181 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     6241 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     6301 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     6361 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     6421 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     6481 c
//
