LOCUS       sequence-368846-        6490 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 6490)
  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 6490)
  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-MPZL2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6490
                     /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(2207..2226)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2239..2257)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2240..2257)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2467..2491)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2488..2691)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2517..2537)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2692..3071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3238..3254
                     /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(3710..3727)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3710..3726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3825..3842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3996..4584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4076..4095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4670..4689)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4695..4714)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4756..4847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4848..4916,4917..5708)
                     /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(5066..5085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5761..5882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5772..5791)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5826..5845
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6012..6490,1..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(6095..6115)
                     /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 gcgccaagga agtggatctc gctgaaccgc acggtgtgca
     1741 ccacagacag tctaatctcg ccgatcacgc cgtccacgtc aggagggttc ttcacttgac
     1801 aggtgtaggt gccgttgtcg tcgaactgca gcttccacag caggatagag gcatcgtatc
     1861 tctcggggtt gccgtcccag gacactctgt ccttgaatct gccgctcata ggctggaagg
     1921 ggtcgatgtg gtagtagaac acgaactgct cagggccgcc atccagaggt ctaaagttcc
     1981 aggtcacggt cagggcatct cccacagggg caaagctgct aaaggtgcac ttcagtctgg
     2041 catcggtgcc gttcacggct tccagcactc tgcttgtgta gatttccacg gcggccgcag
     2101 cgaacaccag gcagaagatg tagctcaggg caatgatggt cttcatggtg gcgctagcag
     2161 gcgcgttttc gatagaaccg aggtgcagtt ggacctggga gtggacacct gtggagagaa
     2221 aggcaaagtg gatgttattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt
     2281 tatgtattaa ttgtagccgc gttctaacga agccaagggg gtgggcctat agactctata
     2341 ggcggtactt acgtcactct tggcacgggg aatccgcgtt ccaatgcacc gttcccggcc
     2401 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
     2461 ccaggcgatc tgacggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2521 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2581 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2641 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2701 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2761 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2821 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2881 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2941 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     3001 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     3061 taatcaatgc caacatggcg gtcatattgg acatgagcca atataaatgt acatattatg
     3121 atatagatac aacgtatgca atggccaata gccaatattg atttatgcta tataaccaat
     3181 gaataatatg gctaatggcc aatattgagt caacgccact gtgcatgaag ctcgtaattg
     3241 ttatccgctc acaattaagg gcgacacaaa atttattcta aatgataata aatactgata
     3301 acatcttata gtttgtatta tattttgtat tatcgttgac atgtataatt ttgatatcaa
     3361 aaactgattt tccctttatt attttcgaga tttattttct taattctctt taacaaacta
     3421 gaaatattgt atatacaaaa aatcataaat aatagatgaa tagtttaatt ataggtgttc
     3481 atcaatcgaa aaagcaacgt atcttattta aagtgcgttg cttttttctc atttataagg
     3541 ttaaataatt ctcatatatc aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc
     3601 tctttcccta aactcccccc ataaaaaaac ccgccgaagc gggtttttac gttatttgcg
     3661 gattaacgat tactcgttat cagaaccgcc cagggggccc gagcttaaga ctggccgtcg
     3721 ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc catccgtcag gggccttctg
     3781 cttagtttga tgcctggcag ttccctactc tcgccttccg cttcctcgct cactgactcg
     3841 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4381 gtaggcggtg ctacagagtt cttgaagtgg tgggctaact acggctacac tagaagaaca
     4441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4561 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4621 cagtggaacg acgcgcgcgt aactcacgtt aagggatttt ggtcatgagc aaggggtgtt
     4681 atgagccata ttcaggtata aatgggctcg cgataatgtt cagaattggt taattggttg
     4741 taacactgac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4801 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatatg agtattcaac
     4861 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4921 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4981 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5041 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5101 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5161 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5221 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5281 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5341 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcgatgg
     5401 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5461 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5521 ctggctggtt tattgctgat aaatccggag ccggtgagcg tggttctcgc ggtatcatcg
     5581 cagcgctggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5701 attggtaatg accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga
     5761 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5821 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5881 ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     5941 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     6001 cgtagcacgt gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     6061 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     6121 gcggatgtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     6181 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     6241 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     6301 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     6361 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     6421 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     6481 tcgatgtggc
//
