LOCUS       sequence-444411-        6760 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 6760)
  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 6760)
  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-SLAMF9-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..123
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     129..148
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      234..822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     723..742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     976..993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1091..1108
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1092..1108
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1564..1580
                     /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        1747..2126
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2127..2330
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2281..2301
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2327..2351
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2561..2579
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2561..2578
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2592..2611
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4082..4126
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4127..4144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4284..4301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4290..4401
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4519..4539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4524..4853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4704..4839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4766..4785
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4901..4948
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4967..5566
                     /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(4967..4986)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5463..5483
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5696..5817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5733..5752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5787..5806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5870..6661,6662..6730))
                     /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     6493..6512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6731..6760,1..62))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
       61 aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat
      121 acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac gtgagttacg
      181 cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      361 gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca cttcaagaac
      421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaggcgagag tagggaactg
     1021 ccaggcatca aactaagcag aaggcccctg acggatggcc tttttgcgtt tctacaaact
     1081 ctttctgtgt tgtaaaacga cggccagtct taagctcggg ccccctgggc ggttctgata
     1141 acgagtaatc gttaatccgc aaataacgta aaaacccgct tcggcgggtt tttttatggg
     1201 gggagtttag ggaaagagca tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat
     1261 atatgagaat tatttaacct tataaatgag aaaaaagcaa cgcactttaa ataagatacg
     1321 ttgctttttc gattgatgaa cacctataat taaactattc atctattatt tatgattttt
     1381 tgtatataca atatttctag tttgttaaag agaattaaga aaataaatct cgaaaataat
     1441 aaagggaaaa tcagtttttg atatcaaaat tatacatgtc aacgataata caaaatataa
     1501 tacaaactat aagatgttat cagtatttat tatcatttag aataaatttt gtgtcgccct
     1561 taattgtgag cggataacaa ttacgagctt catgcacagt ggcgttgact caatattggc
     1621 cattagccat attattcatt ggttatatag cataaatcaa tattggctat tggccattgc
     1681 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     1741 catgttggca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     1801 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     1861 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     1921 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     1981 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     2041 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     2101 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     2161 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     2221 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     2281 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     2341 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
     2401 accgatccag cctccgcggc cgggaacggt gcattggaac gcggattccc cgtgccaaga
     2461 gtgacgtaag taccgcctat agagtctata ggcccacccc cttggcttcg ttagaacgcg
     2521 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
     2581 taacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac tgcacctcgg
     2641 ttctatcgaa aacgcgcctg ctagcgccac catgaagacc atcattgccc tgagctacat
     2701 cttctgcctg gtgttcgctg cggccgccag acgactttgg cgttggtgcg gatctgaaga
     2761 ggtggtggcc gtgctgcaag agagcatcag cctgcctctg gaaatccctc ctgacgagga
     2821 agtggaaaac atcatctggt ccagccacaa gagcctggcc acagtggtgc ctggaaaaga
     2881 aggacaccct gccaccatca tggtcaccaa tcctcactac cagggccaag tgtcctttct
     2941 ggaccctagc tacagcctgc acatcagcaa cctgagctgg gaagatagcg gcctgtatca
     3001 ggcccaagtg aacctgagaa ccagccagat cagcaccatg cagcagtaca acatctgcgt
     3061 gtacagatgg ctgagcgagc cccagatcac cgtgaatttt gagtctagcg gcgagggcgc
     3121 ctgtagcatg tctctcgtgt gttctgtgga aaaggccggc atggacatga cctactcttg
     3181 gctgagcaga ggcgacagca cctacacatt tcacgagggc cctgtgctga gcacatcttg
     3241 gaggcctggc gatagcgccc tgagctacac ctgtagagcc aacaatccca tctccaacgt
     3301 gtccagctgt cccattcctg acggcccttt ctacgccgat cctaactacg ccagcgagaa
     3361 gcctagcacc gccttttgcc ttggcgcgcc gggcggcgac aagacccaca cctgcccccc
     3421 ctgcccagct ccagaactgc tgggagggcc ttccgtgttc ctgttccccc ccaagcccaa
     3481 ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg ccgtgtccca
     3541 cgaggaccca gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa
     3601 gaccaagccc agagaggagc agtacaacag cacctacagg gtggtgtccg tgctgaccgt
     3661 gctgcaccag gactggctga acggcaaaga atacaagtgc aaagtctcca acaaggccct
     3721 ggctgcccca atcgaaaaga caatcagcaa ggccaagggc cagccacggg agccccaggt
     3781 gtacaccctg ccccccagcc gggaggagat gaccaagaac caggtgtccc tgacctgtct
     3841 ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagagcaacg gccagcccga
     3901 gaacaactac aagaccaccc ccccagtgct ggacagcgac ggcagcttct tcctgtacag
     3961 caagctgacc gtggacaagt ccaggtggca gcagggcaac gtgttcagct gcagcgtgat
     4021 gcacgaggcc ctgcacaacc actacaccca gaagtccctg agcctgagcc ccggcggcag
     4081 cggtcttaat gatattttcg aggctcagaa gattgagtgg catgaacacc accaccatca
     4141 ccactaaggc cggcctgtag tcactggcac aacgcgttac ccggtaagcc aatcgggtat
     4201 acacggtcgt catactgcag acagggttct tctactttgc aagatagtct tgagtagtaa
     4261 aataaataga tagagaaaat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4321 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4381 ataaaatgag gaaattgcat cacaacactc aaccctatct cggtctattc ttttgattta
     4441 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4501 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4561 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4621 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4861 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4921 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga ccgagtacaa
     4981 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     5041 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     5101 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5161 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5221 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5281 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5341 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5401 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga catccgcgcc
     5461 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5521 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg tgctacgaga
     5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5701 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5761 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5821 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat taccaatgct
     5881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
     6001 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     6061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6181 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6721 gaatactcat attcttcctt tttcaatatt attgaagcat
//
