LOCUS       sequence-368603-        6517 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 6517)
  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 6517)
  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-SCN3B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     275..295
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    413..524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     513..530
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(670..687)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(688..732)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1960..1979)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1992..2010)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1993..2010)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2220..2244)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2241..2444)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2270..2290)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2445..2824
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2991..3007
                     /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(3463..3480)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3463..3479)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3578..3595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3749..4337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3829..3848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4423..4442)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4448..4467)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4509..4600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4601..4669,4670..5461)
                     /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(4819..4838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5514..5635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5525..5544)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5579..5598
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5765..6364)
                     /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(5848..5868)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6345..6364
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6383..6430)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6478..6517,1..290))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6492..6517,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
       61 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
      121 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      181 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      241 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
      301 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      361 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gtgatgcaat
      421 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      481 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgattttct
      541 ctatctattt attttactac tcaagactat cttgcaaagt agaagaaccc tgtctgcagt
      601 atgacgaccg tgtatacccg attggcttac cgggtaacgc gttgtgccag tgactacagg
      661 ccggccttag tggtgatggt ggtggtgttc atgccactca atcttctgag cctcgaaaat
      721 atcattaaga ccgctgccgc cggggctcag gctcagggac ttctgggtgt agtggttgtg
      781 cagggcctcg tgcatcacgc tgcagctgaa cacgttgccc tgctgccacc tggacttgtc
      841 cacggtcagc ttgctgtaca ggaagaagct gccgtcgctg tccagcactg ggggggtggt
      901 cttgtagttg ttctcgggct ggccgttgct ctcccactcc acggcgatat cgctggggta
      961 gaagcccttc accagacagg tcagggacac ctggttcttg gtcatctcct cccggctggg
     1021 gggcagggtg tacacctggg gctcccgtgg ctggcccttg gccttgctga ttgtcttttc
     1081 gattggggca gccagggcct tgttggagac tttgcacttg tattctttgc cgttcagcca
     1141 gtcctggtgc agcacggtca gcacggacac caccctgtag gtgctgttgt actgctcctc
     1201 tctgggcttg gtcttggcgt tgtgcacctc cacgccgtcc acgtaccagt tgaacttcac
     1261 ctctgggtcc tcgtgggaca cggccaccac cacgcaggtc acctcggggg tcctgctgat
     1321 catcagggtg tccttgggct tgggggggaa caggaacacg gaaggccctc ccagcagttc
     1381 tggagctggg cagggggggc aggtgtgggt cttgtcgccg cccggcgcgc cttcagacac
     1441 cacgctggtg aaatcctcgc cggcttcctc ggtcactctc agagggatca gtctggtggt
     1501 tttcacgaaa ggccggtggg cctcgaactc gaactctctg gacacgttac aggtgtacag
     1561 gccgctgtcg ttcagggtca cgttcagcac ggtgatggac acgtcctgca gatccttaga
     1621 gccgttccac tgcagtctgc cctgaaaggg gctttccacc tcttggtggc cgtttctgta
     1681 ctcgtagatc aggaagtcct tgccgccttc gggtctgtag aaccattcca ccacggttgt
     1741 ggcttccact tcctcgcgct tcatgcagct gatgcatctc agcttcatag ggttgccctg
     1801 cacggcctct gtctcagaag gcacttccac acacactgga aaggcggccg cagcgaacac
     1861 caggcagaag atgtagctca gggcaatgat ggtcttcatg gtggcgctag caggcgcgtt
     1921 ttcgatagaa ccgaggtgca gttggacctg ggagtggaca cctgtggaga gaaaggcaaa
     1981 gtggatgtta ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat
     2041 taattgtagc cgcgttctaa cgaagccaag ggggtgggcc tatagactct ataggcggta
     2101 cttacgtcac tcttggcacg gggaatccgc gttccaatgc accgttcccg gccgcggagg
     2161 ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg
     2221 atctgacggt tcactaaacg agctctgctt atatagacct cccaccgtac acgcctaccg
     2281 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     2341 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     2401 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     2461 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     2521 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     2581 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     2641 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     2701 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     2761 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     2821 tgccaacatg gcggtcatat tggacatgag ccaatataaa tgtacatatt atgatataga
     2881 tacaacgtat gcaatggcca atagccaata ttgatttatg ctatataacc aatgaataat
     2941 atggctaatg gccaatattg agtcaacgcc actgtgcatg aagctcgtaa ttgttatccg
     3001 ctcacaatta agggcgacac aaaatttatt ctaaatgata ataaatactg ataacatctt
     3061 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
     3121 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
     3181 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     3241 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     3301 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     3361 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     3421 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     3481 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     3541 tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     3601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4141 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     4201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4381 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcaaggggt gttatgagcc
     4441 atattcaggt ataaatgggc tcgcgataat gttcagaatt ggttaattgg ttgtaacact
     4501 gacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4561 accctgataa atgcttcaat aatattgaaa aaggaagaat atgagtattc aacatttccg
     4621 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4681 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4741 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4801 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4861 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4921 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4981 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5041 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5101 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcga tggcaacaac
     5161 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5221 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5281 gtttattgct gataaatccg gagccggtga gcgtggttct cgcggtatca tcgcagcgct
     5341 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5401 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5461 atgaccatga ttacgccaag ctctagctag aggtcgacgg tatacagaca tgataagata
     5521 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     5581 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt
     5641 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     5701 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtagca
     5761 cgtgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
     5821 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggatg
     5881 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
     5941 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
     6001 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
     6061 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
     6121 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
     6181 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
     6241 ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
     6301 tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
     6361 tcatggttta gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt
     6421 aattgtcaac acgtgctgat cagatccgaa aatggatata caagctcccg ggagcttttt
     6481 gcaaaagcct aggcctccaa aaaagcctcc tcactac
//
