LOCUS       sequence-420986-        7141 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 7141)
  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 7141)
  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-AMIGO1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(127..147)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        132..461
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      312..447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     374..393
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        509..556
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             575..1174
                     /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(575..594)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1071..1091
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1304..1425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1341..1360)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1395..1414
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1478..2269,2270..2338))
                     /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     2101..2120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2339..2430)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2472..2491
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2497..2516
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2602..3190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3091..3110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3344..3361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3459..3476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3460..3476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3932..3948
                     /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)."
     enhancer        4115..4494
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4495..4698
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4649..4669
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4695..4719
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4929..4947
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4929..4946
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4960..4979
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6831..6875
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6876..6893
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7033..7050)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(7039..7141,1..9)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aattgcatca caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
       61 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
      121 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
      181 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
      241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
      301 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
      361 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
      421 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
      481 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
      541 catagtataa tacgacaagg tgaggaacta aaccatgacc gagtacaagc ccacggtgcg
      601 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
      661 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
      721 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
      781 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
      841 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
      901 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
      961 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     1021 ggcggccgag cgcgccgggg tgcccgcctt cctggagaca tccgcgcccc gcaacctccc
     1081 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     1141 cacctggtgc atgacccgca agcccggtgc ctgacacgtg ctacgagatt tcgattccac
     1201 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1261 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     1321 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1381 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1441 gtcgacctct agctagagct tggcgtaatc atggtcatta ccaatgctta atcagtgagg
     1501 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1561 agataactac gatacgggag ggcttaccat ctggccccag cgctgcgatg ataccgcgag
     1621 aaccacgctc accggctccg gatttatcag caataaacca gccagccgga agggccgagc
     1681 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1741 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc gctacaggca
     1801 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1861 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1921 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1981 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2041 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2101 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2161 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2221 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2281 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatat
     2341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2401 tatttgaatg tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa
     2461 ttctgaacat tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgctca
     2521 tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt
     2581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2761 gccgtagtta gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3361 ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac
     3421 ggatggcctt tttgcgtttc tacaaactct ttctgtgttg taaaacgacg gccagtctta
     3481 agctcgggcc ccctgggcgg ttctgataac gagtaatcgt taatccgcaa ataacgtaaa
     3541 aacccgcttc ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcagaata
     3601 tttaagggcg cctgtcactt tgcttgatat atgagaatta tttaacctta taaatgagaa
     3661 aaaagcaacg cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta
     3721 aactattcat ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag
     3781 aattaagaaa ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta
     3841 tacatgtcaa cgataataca aaatataata caaactataa gatgttatca gtatttatta
     3901 tcatttagaa taaattttgt gtcgccctta attgtgagcg gataacaatt acgagcttca
     3961 tgcacagtgg cgttgactca atattggcca ttagccatat tattcattgg ttatatagca
     4021 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     4081 tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa
     4141 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4201 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4261 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     4321 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     4381 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     4441 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     4501 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     4561 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4621 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     4681 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     4741 tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg ggaacggtgc
     4801 attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag agtctatagg
     4861 cccaccccct tggcttcgtt agaacgcggc tacaattaat acataacctt atgtatcata
     4921 cacatacgat ttaggtgaca ctatagaata acatccactt tgcctttctc tccacaggtg
     4981 tccactccca ggtccaactg cacctcggtt ctatcgaaaa cgcgcctgct agcgccacca
     5041 tgaagaccat cattgccctg agctacatct tctgcctggt gttcgctgcg gccgccggta
     5101 gagctgttgt ttcttgtcct gctgcctgcc tgtgcgccag caatatcctg agctgtagca
     5161 agcagcagct gcccaatgtg cctcacagcc tgccatctta cacagccctg ctggacctga
     5221 gccacaacaa cctgagcaga ctgagagccg agtggacccc tacaagactg acacagctgc
     5281 actccctgct gctgtcccac aatcacctga acttcatcag cagcgaggcc ttctctcccg
     5341 tgcctaacct gagatacctg gacctgtcca gcaaccagct gagaaccctg gacgagttcc
     5401 tgttcagcga cctccaggtg ctggaagtgc tgctgctcta caacaaccac atcatggccg
     5461 tggacagatg cgccttcgat gatatggccc agctgcagaa gctgtatctg agccagaacc
     5521 agatcagcag attcccactg gaactggtca aagagggcgc caagctgcct aagctgacac
     5581 tgctggatct gtcctccaac aagctgaaga atctgcccct gcctgacctg cagaaactgc
     5641 ccgcctggat caagaatggc ctgtacctgc acaacaatcc tctgaactgc gactgcgagc
     5701 tgtaccagct gttcagccac tggcagtaca gacagctgag cagcgtgatg gacttccaag
     5761 aggacctgta ctgcatgaac agcaagaaac tgcacaacgt gttcaacctg tccttcctga
     5821 attgcggcga gtacaaagag agagcctggg aagcccacct gggcgacaca ctgatcatca
     5881 agtgcgacac caaacagcag ggcatgacca aagtgtgggt cacccctagc aacgagcgcg
     5941 tgctggatga agtgaccaat ggcaccgtgt ccgtgtccaa ggatggcagc cttctgttcc
     6001 agcaagtgca ggttgaggac ggcggcgtgt acacatgtta tgccatgggc gagacattca
     6061 acgagacact gagcgtggaa ctgaaggtgc acaacttcac actgcacggc caccacgaca
     6121 ccctgaacac aggcgcgccg ggcggcgaca agacccacac ctgccccccc tgcccagctc
     6181 cagaactgct gggagggcct tccgtgttcc tgttcccccc caagcccaag gacaccctga
     6241 tgatcagcag gacccccgag gtgacctgcg tggtggtggc cgtgtcccac gaggacccag
     6301 aggtgaagtt caactggtac gtggacggcg tggaggtgca caacgccaag accaagccca
     6361 gagaggagca gtacaacagc acctacaggg tggtgtccgt gctgaccgtg ctgcaccagg
     6421 actggctgaa cggcaaagaa tacaagtgca aagtctccaa caaggccctg gctgccccaa
     6481 tcgaaaagac aatcagcaag gccaagggcc agccacggga gccccaggtg tacaccctgc
     6541 cccccagccg ggaggagatg accaagaacc aggtgtccct gacctgtctg gtgaagggct
     6601 tctaccccag cgatatcgcc gtggagtggg agagcaacgg ccagcccgag aacaactaca
     6661 agaccacccc cccagtgctg gacagcgacg gcagcttctt cctgtacagc aagctgaccg
     6721 tggacaagtc caggtggcag cagggcaacg tgttcagctg cagcgtgatg cacgaggccc
     6781 tgcacaacca ctacacccag aagtccctga gcctgagccc cggcggcagc ggtcttaatg
     6841 atattttcga ggctcagaag attgagtggc atgaacacca ccaccatcac cactaaggcc
     6901 ggcctgtagt cactggcaca acgcgttacc cggtaagcca atcgggtata cacggtcgtc
     6961 atactgcaga cagggttctt ctactttgca agatagtctt gagtagtaaa ataaatagat
     7021 agagaaaatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     7081 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     7141 a
//
