LOCUS       sequence-386082-        7027 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 7027)
  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 7027)
  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-ZPBP-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7027
                     /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             2603..2647
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2648..2665
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2805..2822)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2811..2922
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3040..3060)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3045..3374
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3225..3360
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3287..3306
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3422..3469
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3488..4087
                     /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     3984..4004
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4254..4273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4308..4327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4391..5182,5183..5251))
                     /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     5014..5033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5252..5343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5385..5404
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5410..5429
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5515..6103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6004..6023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6257..6274
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6372..6389
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6373..6389
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6845..6861
                     /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 cccatcttgt tagactgcct agagccttcc ggctgaccaa
     1021 ggactctgtg aagatcgtgg gcagcaccag ctttcccgtg aaggcctatg tgatgctgca
     1081 ccagaaaagc cctcacgtgc tgtgtgtgac ccagcagctg agaaacgccg agctgatcga
     1141 tcccagcttt cagtggtacg gccccaaggg caaagtggtg tccgtggaaa acagaacagc
     1201 ccagatcacc agcaccggca gcctggtgtt ccagaacttc gaggaatcca tgagcgggat
     1261 ctatacctgc ttcctcgagt acaagcccac cgtggaagag atcgtgaagc ggctgcagct
     1321 gaagtacgcc atctacgcct acagagagcc ccactactac taccagttca ccgccagata
     1381 ccacgctgcc ccttgcaaca gcatctacaa catcagcttc gagaagaagc tgctgcagat
     1441 cctgagcaaa ctgctgctgg acctgagctg cgagatcagc ctgctgaagt ccgagtgcca
     1501 cagagtgaag atgcagagag ccggactgca gaacgagctg ttcttcgcct ttagcgtgtc
     1561 cagcctggat accgagaagg gccctaagag atgcaccgac cacaactgcg agccctacaa
     1621 gagactgttc aaggccaaga acctgatcga gcggttcttc aaccagcaag tggaaatcct
     1681 cggcagaagg gccgaacagc tgccccagat ctactacatc gagggcacac tgcagatggt
     1741 ctggatcaac agatgcttcc ccggctacgg catgaacgtg cagcagcacc ctaagtgccc
     1801 tgagtgttgc gtgatctgta gccccggcag ctacaaccct agagatggca ttcactgcct
     1861 gcagtgcaac agctctctgg tgtacggcgc caagacctgc cttggcgcgc cgggcggcga
     1921 caagacccac acctgccccc cctgcccagc tccagaactg ctgggagggc cttccgtgtt
     1981 cctgttcccc cccaagccca aggacaccct gatgatcagc aggacccccg aggtgacctg
     2041 cgtggtggtg gccgtgtccc acgaggaccc agaggtgaag ttcaactggt acgtggacgg
     2101 cgtggaggtg cacaacgcca agaccaagcc cagagaggag cagtacaaca gcacctacag
     2161 ggtggtgtcc gtgctgaccg tgctgcacca ggactggctg aacggcaaag aatacaagtg
     2221 caaagtctcc aacaaggccc tggctgcccc aatcgaaaag acaatcagca aggccaaggg
     2281 ccagccacgg gagccccagg tgtacaccct gccccccagc cgggaggaga tgaccaagaa
     2341 ccaggtgtcc ctgacctgtc tggtgaaggg cttctacccc agcgatatcg ccgtggagtg
     2401 ggagagcaac ggccagcccg agaacaacta caagaccacc cccccagtgc tggacagcga
     2461 cggcagcttc ttcctgtaca gcaagctgac cgtggacaag tccaggtggc agcagggcaa
     2521 cgtgttcagc tgcagcgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct
     2581 gagcctgagc cccggcggca gcggtcttaa tgatattttc gaggctcaga agattgagtg
     2641 gcatgaacac caccaccatc accactaagg ccggcctgta gtcactggca caacgcgtta
     2701 cccggtaagc caatcgggta tacacggtcg tcatactgca gacagggttc ttctactttg
     2761 caagatagtc ttgagtagta aaataaatag atagagaaaa tcagcctcga ctgtgccttc
     2821 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2881 cactcccact gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc
     2941 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3001 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3121 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3181 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3241 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3301 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3361 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     3421 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3481 ctaaaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     3541 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3601 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3661 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3721 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     3781 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     3841 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     3901 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     3961 cttcctggag acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4021 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4081 tgcctgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4141 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4201 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4261 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4321 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4381 atcatggtca ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4501 catctggccc cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat
     4561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4681 gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5041 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5221 taagggcgac acggaaatgt tgaatactca tattcttcct ttttcaatat tattgaagca
     5281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5341 aaataggggt cagtgttaca accaattaac caattctgaa cattatcgcg agcccattta
     5401 tacctgaata tggctcataa caccccttgc tcatgaccaa aatcccttaa cgtgagttac
     5461 gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5521 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5581 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5641 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttagcccacc acttcaagaa
     5701 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5761 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5821 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5881 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5941 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6001 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6061 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6121 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6181 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6241 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaggcgaga gtagggaact
     6301 gccaggcatc aaactaagca gaaggcccct gacggatggc ctttttgcgt ttctacaaac
     6361 tctttctgtg ttgtaaaacg acggccagtc ttaagctcgg gccccctggg cggttctgat
     6421 aacgagtaat cgttaatccg caaataacgt aaaaacccgc ttcggcgggt ttttttatgg
     6481 ggggagttta gggaaagagc atttgtcaga atatttaagg gcgcctgtca ctttgcttga
     6541 tatatgagaa ttatttaacc ttataaatga gaaaaaagca acgcacttta aataagatac
     6601 gttgcttttt cgattgatga acacctataa ttaaactatt catctattat ttatgatttt
     6661 ttgtatatac aatatttcta gtttgttaaa gagaattaag aaaataaatc tcgaaaataa
     6721 taaagggaaa atcagttttt gatatcaaaa ttatacatgt caacgataat acaaaatata
     6781 atacaaacta taagatgtta tcagtattta ttatcattta gaataaattt tgtgtcgccc
     6841 ttaattgtga gcggataaca attacgagct tcatgcacag tggcgttgac tcaatattgg
     6901 ccattagcca tattattcat tggttatata gcataaatca atattggcta ttggccattg
     6961 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     7021 ccatgtt
//
