LOCUS       sequence-368531-        6595 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 6595)
  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 6595)
  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-CD300A-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     443..463
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    676..797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(713..732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     767..786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(850..1641,1642..1710))
                     /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     1473..1492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1711..1802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1844..1863
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1869..1888
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1974..2562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2463..2482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2716..2733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2831..2848
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2832..2848
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3304..3320
                     /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        3487..3866
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3867..4070
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4021..4041
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4067..4091
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4301..4319
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4301..4318
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4332..4351
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5657..5701
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5702..5719
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5859..5876)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5865..5976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6094..6114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6099..6428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6279..6414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6341..6360
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6476..6523
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(6542..6595,1..546)
                     /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(6542..6561)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
       61 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
      121 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
      181 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
      241 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
      301 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
      361 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
      421 ttcctggaga catccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
      481 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
      541 gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
      601 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
      661 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
      721 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
      781 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
      841 tcatggtcat taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      961 atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc cggatttatc
     1021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1141 tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1681 aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt attgaagcat
     1741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1801 aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat
     1861 acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac gtgagttacg
     1921 cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2101 gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca cttcaagaac
     2161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2701 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaggcgagag tagggaactg
     2761 ccaggcatca aactaagcag aaggcccctg acggatggcc tttttgcgtt tctacaaact
     2821 ctttctgtgt tgtaaaacga cggccagtct taagctcggg ccccctgggc ggttctgata
     2881 acgagtaatc gttaatccgc aaataacgta aaaacccgct tcggcgggtt tttttatggg
     2941 gggagtttag ggaaagagca tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat
     3001 atatgagaat tatttaacct tataaatgag aaaaaagcaa cgcactttaa ataagatacg
     3061 ttgctttttc gattgatgaa cacctataat taaactattc atctattatt tatgattttt
     3121 tgtatataca atatttctag tttgttaaag agaattaaga aaataaatct cgaaaataat
     3181 aaagggaaaa tcagtttttg atatcaaaat tatacatgtc aacgataata caaaatataa
     3241 tacaaactat aagatgttat cagtatttat tatcatttag aataaatttt gtgtcgccct
     3301 taattgtgag cggataacaa ttacgagctt catgcacagt ggcgttgact caatattggc
     3361 cattagccat attattcatt ggttatatag cataaatcaa tattggctat tggccattgc
     3421 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     3481 catgttggca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     3541 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3601 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3661 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3721 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3781 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     3841 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     3901 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     3961 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4021 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     4081 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
     4141 accgatccag cctccgcggc cgggaacggt gcattggaac gcggattccc cgtgccaaga
     4201 gtgacgtaag taccgcctat agagtctata ggcccacccc cttggcttcg ttagaacgcg
     4261 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
     4321 taacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac tgcacctcgg
     4381 ttctatcgaa aacgcgcctg ctagcgccac catgaagacc atcattgccc tgagctacat
     4441 cttctgcctg gtgttcgctg cggccgcctt gagcaagtgt agaacagtgg ctggccctgt
     4501 cggcggatct ctgtctgttc agtgccccta cgagaaagag cacagaaccc tgaacaagta
     4561 ctggtgccgg cctcctcaga tcttcctgtg cgacaagatc gtggaaacaa agggcagcgc
     4621 cggcaagaga aacggcagag tgtctatcag agacagcccc gccaacctga gcttcaccgt
     4681 gacactggaa aacctgaccg aggaagatgc cggcacatat tggtgcggag tggacacccc
     4741 ttggctgaga gactttcacg accccgtggt ggaagtggaa gtgtccgtgt ttcctgccag
     4801 caccagcatg acaccagcct ctattacagc cgccaagacc agcaccatca ccaccgcttt
     4861 tccacctgtg tccagcacca cactgtttgc cgtgggagcc acacacagcg ccagcatcca
     4921 agaagagaca gaggaagtcg tgaacagcca gctgcctggc gcgccgggcg gcgacaagac
     4981 ccacacctgc cccccctgcc cagctccaga actgctggga gggccttccg tgttcctgtt
     5041 cccccccaag cccaaggaca ccctgatgat cagcaggacc cccgaggtga cctgcgtggt
     5101 ggtggccgtg tcccacgagg acccagaggt gaagttcaac tggtacgtgg acggcgtgga
     5161 ggtgcacaac gccaagacca agcccagaga ggagcagtac aacagcacct acagggtggt
     5221 gtccgtgctg accgtgctgc accaggactg gctgaacggc aaagaataca agtgcaaagt
     5281 ctccaacaag gccctggctg ccccaatcga aaagacaatc agcaaggcca agggccagcc
     5341 acgggagccc caggtgtaca ccctgccccc cagccgggag gagatgacca agaaccaggt
     5401 gtccctgacc tgtctggtga agggcttcta ccccagcgat atcgccgtgg agtgggagag
     5461 caacggccag cccgagaaca actacaagac caccccccca gtgctggaca gcgacggcag
     5521 cttcttcctg tacagcaagc tgaccgtgga caagtccagg tggcagcagg gcaacgtgtt
     5581 cagctgcagc gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct
     5641 gagccccggc ggcagcggtc ttaatgatat tttcgaggct cagaagattg agtggcatga
     5701 acaccaccac catcaccact aaggccggcc tgtagtcact ggcacaacgc gttacccggt
     5761 aagccaatcg ggtatacacg gtcgtcatac tgcagacagg gttcttctac tttgcaagat
     5821 agtcttgagt agtaaaataa atagatagag aaaatcagcc tcgactgtgc cttctagttg
     5881 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     5941 cactgtcctt tcctaataaa atgaggaaat tgcatcacaa cactcaaccc tatctcggtc
     6001 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6061 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     6121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6361 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6421 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     6481 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     6541 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagg
//
