LOCUS       sequence-425640-        7615 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 7615)
  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 7615)
  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-DLL4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3191..3235
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3236..3253
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3393..3410)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3399..3510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3628..3648)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3633..3962
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3813..3948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3875..3894
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4010..4057
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4076..4675
                     /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(4076..4095)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4572..4592
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4805..4926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4842..4861)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4896..4915
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4979..5770,5771..5839))
                     /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     5602..5621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5840..5931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5973..5992
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5998..6017
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6103..6691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6592..6611
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6845..6862
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6960..6977
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6961..6977
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7433..7449
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg ccagcggagt ttttcagctg caactgcaag agttcatcaa
     1021 cgagagaggc gtgctggcct ctggcagacc ttgtgaacct ggctgccgga ccttcttcag
     1081 agtgtgcctg aagcacttcc aggccgtggt gtctcctgga ccttgtacct ttggcaccgt
     1141 gtctacacct gtgctgggca ccaatagctt cgccgtgcgg gatgattcta gcggcggagg
     1201 cagaaatcct ctgcagctgc ccttcaactt cacctggcct ggcaccttca gcctgatcat
     1261 cgaagcctgg catgcccctg gggatgatct gaggcctgaa gctttgcctc ctgacgctct
     1321 gatcagcaag atcgccatcc agggatctct ggccgtgggc caaaattggc tgctggacga
     1381 gcagaccagc acactgacca gactgcggta cagctacaga gtgatctgca gcgacaacta
     1441 ctacggcgac aactgcagca gactgtgcaa gaagcggaac gaccacttcg gccactacgt
     1501 gtgtcagccc gatggcaacc tgtcttgtct gcctggatgg accggcgagt actgccagca
     1561 acctatctgt ctgagcggct gccacgagca gaatggctac tgttctaagc ccgccgagtg
     1621 cctgtgtaga cctggctggc aaggcaggct gtgcaacgag tgcatccctc acaatggctg
     1681 cagacacggc acctgtagca ccccttggca gtgcacatgt gacgaaggat ggggcggact
     1741 gttctgcgac caagacctga actactgcac ccatcacagc ccctgcaaga acggcgccac
     1801 atgtagcaac agcggccagc ggtcttacac ctgtacctgc agacctggat acaccggcgt
     1861 ggactgcgag ctggaactga gcgagtgcga cagcaacccc tgtagaaatg gcggctcctg
     1921 caaggaccaa gaggacggct atcactgcct gtgtccacct ggatattacg gcctgcactg
     1981 cgaacacagc accctgagct gtgccgactc tccctgtttt aatggcggca gctgtagaga
     2041 gagaaaccag ggcgccaatt acgcctgcga gtgcccacct aatttcaccg gcagcaactg
     2101 cgagaagaaa gtggaccggt gcaccagcaa tccctgcgct aatggtggcc agtgcctgaa
     2161 cagaggcccc agcagaatgt gtagatgcag acccggcttt accggcacct attgcgaact
     2221 gcacgtgtcc gactgcgccc ggaatccttg tgctcatggc ggcacatgcc acgacctgga
     2281 aaacggcctg atgtgcactt gccctgccgg cttctctggg agaagatgtg aagtgcggac
     2341 cagcatcgac gcctgtgcca gctctccttg cttcaacaga gccacctgtt acaccgacct
     2401 gagcaccgac accttcgtgt gcaactgccc ttacggcttc gtgggcagca gatgcgagtt
     2461 tcctgttggc ctgcctccta gcttcccttg gggcgcgccg ggcggcgaca agacccacac
     2521 ctgccccccc tgcccagctc cagaactgct gggagggcct tccgtgttcc tgttcccccc
     2581 caagcccaag gacaccctga tgatcagcag gacccccgag gtgacctgcg tggtggtggc
     2641 cgtgtcccac gaggacccag aggtgaagtt caactggtac gtggacggcg tggaggtgca
     2701 caacgccaag accaagccca gagaggagca gtacaacagc acctacaggg tggtgtccgt
     2761 gctgaccgtg ctgcaccagg actggctgaa cggcaaagaa tacaagtgca aagtctccaa
     2821 caaggccctg gctgccccaa tcgaaaagac aatcagcaag gccaagggcc agccacggga
     2881 gccccaggtg tacaccctgc cccccagccg ggaggagatg accaagaacc aggtgtccct
     2941 gacctgtctg gtgaagggct tctaccccag cgatatcgcc gtggagtggg agagcaacgg
     3001 ccagcccgag aacaactaca agaccacccc cccagtgctg gacagcgacg gcagcttctt
     3061 cctgtacagc aagctgaccg tggacaagtc caggtggcag cagggcaacg tgttcagctg
     3121 cagcgtgatg cacgaggccc tgcacaacca ctacacccag aagtccctga gcctgagccc
     3181 cggcggcagc ggtcttaatg atattttcga ggctcagaag attgagtggc atgaacacca
     3241 ccaccatcac cactaaggcc ggcctgtagt cactggcaca acgcgttacc cggtaagcca
     3301 atcgggtata cacggtcgtc atactgcaga cagggttctt ctactttgca agatagtctt
     3361 gagtagtaaa ataaatagat agagaaaatc agcctcgact gtgccttcta gttgccagcc
     3421 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3481 cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct
     3541 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3601 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3661 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3721 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3781 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3841 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3901 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3961 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     4021 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac
     4081 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4141 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4201 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4261 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4321 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4381 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4441 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4501 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4561 atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4621 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt
     4681 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4741 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4801 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4861 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4921 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt
     4981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5101 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
     5161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5281 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5821 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
     5941 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     6001 gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     6061 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6121 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6181 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6241 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     6301 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6361 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6421 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6481 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6541 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6601 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6661 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6721 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6781 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6841 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     6901 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     6961 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     7021 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     7081 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     7141 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     7201 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     7261 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     7321 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     7381 agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt aattgtgagc
     7441 ggataacaat tacgagcttc atgcacagtg gcgttgactc aatattggcc attagccata
     7501 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7561 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
