LOCUS       sequence-368840-        6487 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 6487)
  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 6487)
  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-MPZL1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(140..187)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(235..564)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(303..322)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     549..569
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    687..798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     787..804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(944..961)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(962..1006)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2204..2223)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2236..2254)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2237..2254)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2464..2488)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2485..2688)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2514..2534)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2689..3068
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3235..3251
                     /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(3707..3724)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3707..3723)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3822..3839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3993..4581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4073..4092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4667..4686)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4692..4711)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4753..4844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4845..4913,4914..5705)
                     /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(5063..5082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5758..5879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5769..5788)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5823..5842
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6009..6487,1..121))
                     /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(6092..6112)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
       61 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
      121 tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat
      181 tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
      241 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc
      301 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
      361 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      421 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      481 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      541 ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa
      601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      661 tagaccgaga tagggttgag tgttgtgatg caatttcctc attttattag gaaaggacag
      721 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      781 ggcaactaga aggcacagtc gaggctgatt ttctctatct atttatttta ctactcaaga
      841 ctatcttgca aagtagaaga accctgtctg cagtatgacg accgtgtata cccgattggc
      901 ttaccgggta acgcgttgtg ccagtgacta caggccggcc ttagtggtga tggtggtggt
      961 gttcatgcca ctcaatcttc tgagcctcga aaatatcatt aagaccgctg ccgccggggc
     1021 tcaggctcag ggacttctgg gtgtagtggt tgtgcagggc ctcgtgcatc acgctgcagc
     1081 tgaacacgtt gccctgctgc cacctggact tgtccacggt cagcttgctg tacaggaaga
     1141 agctgccgtc gctgtccagc actggggggg tggtcttgta gttgttctcg ggctggccgt
     1201 tgctctccca ctccacggcg atatcgctgg ggtagaagcc cttcaccaga caggtcaggg
     1261 acacctggtt cttggtcatc tcctcccggc tggggggcag ggtgtacacc tggggctccc
     1321 gtggctggcc cttggccttg ctgattgtct tttcgattgg ggcagccagg gccttgttgg
     1381 agactttgca cttgtattct ttgccgttca gccagtcctg gtgcagcacg gtcagcacgg
     1441 acaccaccct gtaggtgctg ttgtactgct cctctctggg cttggtcttg gcgttgtgca
     1501 cctccacgcc gtccacgtac cagttgaact tcacctctgg gtcctcgtgg gacacggcca
     1561 ccaccacgca ggtcacctcg ggggtcctgc tgatcatcag ggtgtccttg ggcttggggg
     1621 ggaacaggaa cacggaaggc cctcccagca gttctggagc tgggcagggg gggcaggtgt
     1681 gggtcttgtc gccgcccggc gcgccaacag gcaggttctc tttttccacc acgtacagtc
     1741 tgatgtggcc aggctgcacc acgatgtcag gagggttctt cacgtcgcag atgtaggtcc
     1801 cgttgtggat gaactgcatg ttctcgatgt tgatgctggc gtccttcttg tccaggtcgc
     1861 cagcccagct gattctgtcc ttgaatggag ggtagttgcc caggtacact tggccctggc
     1921 tgtagtgaaa gaagctgacg gttgtatcgg cgccctcagg ctgaaaagac caagacacag
     1981 aggtcaggcc gcctgtggtg ctggtgctct taaacttgca ggtcagcttg ccctgggtgc
     2041 cgttggccac gaagatttct ttaggggtgt acacttccag agcagaggcg gccgcagcga
     2101 acaccaggca gaagatgtag ctcagggcaa tgatggtctt catggtggcg ctagcaggcg
     2161 cgttttcgat agaaccgagg tgcagttgga cctgggagtg gacacctgtg gagagaaagg
     2221 caaagtggat gttattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat
     2281 gtattaattg tagccgcgtt ctaacgaagc caagggggtg ggcctataga ctctataggc
     2341 ggtacttacg tcactcttgg cacggggaat ccgcgttcca atgcaccgtt cccggccgcg
     2401 gaggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca
     2461 ggcgatctga cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct
     2521 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     2581 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     2641 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     2701 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     2761 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     2821 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     2881 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     2941 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     3001 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     3061 tcaatgccaa catggcggtc atattggaca tgagccaata taaatgtaca tattatgata
     3121 tagatacaac gtatgcaatg gccaatagcc aatattgatt tatgctatat aaccaatgaa
     3181 taatatggct aatggccaat attgagtcaa cgccactgtg catgaagctc gtaattgtta
     3241 tccgctcaca attaagggcg acacaaaatt tattctaaat gataataaat actgataaca
     3301 tcttatagtt tgtattatat tttgtattat cgttgacatg tataattttg atatcaaaaa
     3361 ctgattttcc ctttattatt ttcgagattt attttcttaa ttctctttaa caaactagaa
     3421 atattgtata tacaaaaaat cataaataat agatgaatag tttaattata ggtgttcatc
     3481 aatcgaaaaa gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt tataaggtta
     3541 aataattctc atatatcaag caaagtgaca ggcgccctta aatattctga caaatgctct
     3601 ttccctaaac tccccccata aaaaaacccg ccgaagcggg tttttacgtt atttgcggat
     3661 taacgattac tcgttatcag aaccgcccag ggggcccgag cttaagactg gccgtcgttt
     3721 tacaacacag aaagagtttg tagaaacgca aaaaggccat ccgtcagggg ccttctgctt
     3781 agtttgatgc ctggcagttc cctactctcg ccttccgctt cctcgctcac tgactcgctg
     3841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4381 ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag aagaacagta
     4441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4621 tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagcaag gggtgttatg
     4681 agccatattc aggtataaat gggctcgcga taatgttcag aattggttaa ttggttgtaa
     4741 cactgacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4801 aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatatgagt attcaacatt
     4861 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4921 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4981 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5041 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5101 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5161 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5221 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5281 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5341 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcgatggcaa
     5401 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5461 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5521 gctggtttat tgctgataaa tccggagccg gtgagcgtgg ttctcgcggt atcatcgcag
     5581 cgctggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5641 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5701 ggtaatgacc atgattacgc caagctctag ctagaggtcg acggtataca gacatgataa
     5761 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     5821 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
     5881 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     5941 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     6001 agcacgtgtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
     6061 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
     6121 gatgtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
     6181 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
     6241 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
     6301 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
     6361 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
     6421 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
     6481 atgtggc
//
