LOCUS       sequence-405790-        6835 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 6835)
  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 6835)
  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-CD72-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6835
                     /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             2411..2455
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2456..2473
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2613..2630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2619..2730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2848..2868)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2853..3182
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3033..3168
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3095..3114
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3230..3277
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3296..3895
                     /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(3296..3315)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3792..3812
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4025..4146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4062..4081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4116..4135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4199..4990,4991..5059))
                     /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     4822..4841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5060..5151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5193..5212
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5218..5237
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5323..5911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5812..5831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6065..6082
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6180..6197
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6181..6197
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6653..6669
                     /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 ccagatacct gcaagtttct cagcagctgc agcagaccaa
     1021 cagagtgctg gaagtgacca acagcagcct gagacagcag ctgaggctga agatcacaca
     1081 gctgggacag agcgccgagg atctgcaggg aagtagaaga gagctggccc agtctcaaga
     1141 ggccctgcag gttgaacaga gagcccatca agctgccgaa ggacagctgc aggcttgcca
     1201 ggccgaccgg cagaaaacaa aagagacact gcagagcgag gaacagcagc ggagagccct
     1261 ggaacagaaa ctgagcaaca tggaaaaccg gctgaagcca ttcttcacct gtggcagcgc
     1321 cgacacatgt tgtcctagcg gctggatcat gcaccagaag tcctgcttct acatcagcct
     1381 gaccagcaag aactggcaag agtcccagaa gcagtgcgag acactgagca gcaagctggc
     1441 caccttcagc gagatctacc ctcagagcca cagctactac tttctgaaca gcctgctgcc
     1501 taacggcggc agcggcaatt cttactggac cggcctgtct agcaacaagg actggaagct
     1561 gaccgacgac acccagagaa ccagaacata cgcccagtcc agcaagtgca acaaggtgca
     1621 caagacctgg tcctggtgga ccctggaaag cgagagctgt agaagctccc tgccttacat
     1681 ctgcgagatg accgccttca gattccccga tggcgcgccg ggcggcgaca agacccacac
     1741 ctgccccccc tgcccagctc cagaactgct gggagggcct tccgtgttcc tgttcccccc
     1801 caagcccaag gacaccctga tgatcagcag gacccccgag gtgacctgcg tggtggtggc
     1861 cgtgtcccac gaggacccag aggtgaagtt caactggtac gtggacggcg tggaggtgca
     1921 caacgccaag accaagccca gagaggagca gtacaacagc acctacaggg tggtgtccgt
     1981 gctgaccgtg ctgcaccagg actggctgaa cggcaaagaa tacaagtgca aagtctccaa
     2041 caaggccctg gctgccccaa tcgaaaagac aatcagcaag gccaagggcc agccacggga
     2101 gccccaggtg tacaccctgc cccccagccg ggaggagatg accaagaacc aggtgtccct
     2161 gacctgtctg gtgaagggct tctaccccag cgatatcgcc gtggagtggg agagcaacgg
     2221 ccagcccgag aacaactaca agaccacccc cccagtgctg gacagcgacg gcagcttctt
     2281 cctgtacagc aagctgaccg tggacaagtc caggtggcag cagggcaacg tgttcagctg
     2341 cagcgtgatg cacgaggccc tgcacaacca ctacacccag aagtccctga gcctgagccc
     2401 cggcggcagc ggtcttaatg atattttcga ggctcagaag attgagtggc atgaacacca
     2461 ccaccatcac cactaaggcc ggcctgtagt cactggcaca acgcgttacc cggtaagcca
     2521 atcgggtata cacggtcgtc atactgcaga cagggttctt ctactttgca agatagtctt
     2581 gagtagtaaa ataaatagat agagaaaatc agcctcgact gtgccttcta gttgccagcc
     2641 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2701 cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct
     2761 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2821 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3001 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3061 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3121 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3181 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     3241 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac
     3301 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3361 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3421 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3481 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3541 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3601 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3661 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3721 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     3781 atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     3841 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt
     3901 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3961 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4021 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4081 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4141 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt
     4201 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4261 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4321 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
     4381 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4441 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4501 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4561 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4621 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4681 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4741 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4801 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4861 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4921 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4981 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5041 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5101 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
     5161 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     5221 gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     5281 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5341 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5401 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5461 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     5521 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5581 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5641 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5701 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5761 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5821 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5881 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5941 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6001 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6061 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     6121 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     6181 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     6241 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     6301 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     6361 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     6421 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     6481 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     6541 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     6601 agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt aattgtgagc
     6661 ggataacaat tacgagcttc atgcacagtg gcgttgactc aatattggcc attagccata
     6721 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     6781 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
