LOCUS       sequence-368520-        6568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6568)
  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 6568)
  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-TREML4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2290..2309)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2322..2340)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2323..2340)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2550..2574)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2571..2774)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2600..2620)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2775..3154
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3321..3337
                     /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(3793..3810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3793..3809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3908..3925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4079..4667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4159..4178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4753..4772)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4778..4797)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4839..4930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4931..4999,5000..5791)
                     /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(5149..5168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5844..5965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5855..5874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5909..5928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6095..6568,1..126))
                     /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(6178..6198)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc agagccgagg caggcagggg ctctgctttt
     1741 tcttgtctcg ctgccattga tgctggggtg tccgcttgtg ccctcagggc ttgtaatcag
     1801 cactgtgctg gtaggcagcc aaggcagtgt ccacatagga gaggtggtag gagcagggga
     1861 caccaccagg ctgatattcc gcagcacggt gatgatgttc tcgctggcgt tgtagatgcc
     1921 gcaccaatag aagccgctgt cgttctgggt cagctggatc atggtgatgt tgaagaagcc
     1981 ggcgttgggc ttatcccaga tggtgtagtg ggacttctgc acggctgtcc aaggcttaga
     2041 gctggtcacg agcagggtac atctgctagg gcttgtctgc tgacaccagc tcttaggctg
     2101 gtaggggcct ctcttgggag aatactggca ctgcagcagc agggtctgtc caggatgttt
     2161 gtgcagttct tcggcggccg cagcgaacac caggcagaag atgtagctca gggcaatgat
     2221 ggtcttcatg gtggcgctag caggcgcgtt ttcgatagaa ccgaggtgca gttggacctg
     2281 ggagtggaca cctgtggaga gaaaggcaaa gtggatgtta ttctatagtg tcacctaaat
     2341 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgaagccaag
     2401 ggggtgggcc tatagactct ataggcggta cttacgtcac tcttggcacg gggaatccgc
     2461 gttccaatgc accgttcccg gccgcggagg ctggatcggt cccggtgtct tctatggagg
     2521 tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt
     2581 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
     2641 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     2701 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
     2761 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     2821 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     2881 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     2941 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     3001 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     3061 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     3121 gattactatt aataactagt caataatcaa tgccaacatg gcggtcatat tggacatgag
     3181 ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca atagccaata
     3241 ttgatttatg ctatataacc aatgaataat atggctaatg gccaatattg agtcaacgcc
     3301 actgtgcatg aagctcgtaa ttgttatccg ctcacaatta agggcgacac aaaatttatt
     3361 ctaaatgata ataaatactg ataacatctt atagtttgta ttatattttg tattatcgtt
     3421 gacatgtata attttgatat caaaaactga ttttcccttt attattttcg agatttattt
     3481 tcttaattct ctttaacaaa ctagaaatat tgtatataca aaaaatcata aataatagat
     3541 gaatagttta attataggtg ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg
     3601 ttgctttttt ctcatttata aggttaaata attctcatat atcaagcaaa gtgacaggcg
     3661 cccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga
     3721 agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc gcccaggggg
     3781 cccgagctta agactggccg tcgttttaca acacagaaag agtttgtaga aacgcaaaaa
     3841 ggccatccgt caggggcctt ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt
     3901 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3961 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtgggcta
     4501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4681 tcttttctac ggggtctgac gctcagtgga acgacgcgcg cgtaactcac gttaagggat
     4741 tttggtcatg agcaaggggt gttatgagcc atattcaggt ataaatgggc tcgcgataat
     4801 gttcagaatt ggttaattgg ttgtaacact gacccctatt tgtttatttt tctaaataca
     4861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4921 aaggaagaat atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5161 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5461 caccacgatg cctgtagcga tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatccg gagccggtga
     5641 gcgtggttct cgcggtatca tcgcagcgct ggggccagat ggtaagccct cccgtatcgt
     5701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5761 gataggtgcc tcactgatta agcattggta atgaccatga ttacgccaag ctctagctag
     5821 aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5881 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5941 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
     6001 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     6061 aggcggcggt ggaatcgaaa tctcgtagca cgtgtcaggc accgggcttg cgggtcatgc
     6121 accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct
     6181 cgtagaaggg gaggttgcgg ggcgcggatg tctccaggaa ggcgggcacc ccggcgcgct
     6241 cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg
     6301 gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga
     6361 ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc
     6421 cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg
     6481 cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga
     6541 gttcttgcag ctcggtgacc cgctcgat
//
