LOCUS       sequence-424401-        6760 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 6760)
  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 6760)
  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-CXADR-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6760
                     /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             2336..2380
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2381..2398
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2538..2555)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2544..2655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2773..2793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2778..3107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2958..3093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3020..3039
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3155..3202
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3221..3820
                     /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(3221..3240)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3717..3737
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3950..4071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3987..4006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4041..4060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4124..4915,4916..4984))
                     /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     4747..4766
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4985..5076)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5118..5137
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5143..5162
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5248..5836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5737..5756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5990..6007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6105..6122
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6106..6122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6578..6594
                     /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 ccttgagcat caccacacct gaggaaatga tcgagaaggc
     1021 caagggcgag acagcctacc tgccttgcaa gttcacactg agccctgagg atcagggccc
     1081 tctggatatc gagtggctga tctcccctgc cgacaaccag aaagtggacc aagtgatcat
     1141 cctgtacagc ggcgacaaga tctacgacga ctactacccc gacctgaagg gcagagtgca
     1201 cttcaccagc aacgatctga agtccggcga cgccagcatc aacgtgacaa acctgcagct
     1261 gagcgacatc ggcacctacc agtgcaaagt gaagaaagcc cctggcgtgg ccaacaagaa
     1321 aatccacctg gtggtgctgg tcaagcctag cggcgccaga tgttatgtgg acggctctga
     1381 agagatcggc agcgacttca agatcaagtg cgagcccaaa gagggcagcc tgcctctgca
     1441 gtatgagtgg cagaagctgt ccgacagcca gaagatgcct acaagctggc tggccgagat
     1501 gaccagcagc gtgatcagcg tgaagaacgc cagcagcgag tactctggca catacagctg
     1561 caccgtgcgg aacagagtgg gctctgatca gtgcctgctg agactgaatg tggtgcctcc
     1621 tagcaacaag gccggtggcg cgccgggcgg cgacaagacc cacacctgcc ccccctgccc
     1681 agctccagaa ctgctgggag ggccttccgt gttcctgttc ccccccaagc ccaaggacac
     1741 cctgatgatc agcaggaccc ccgaggtgac ctgcgtggtg gtggccgtgt cccacgagga
     1801 cccagaggtg aagttcaact ggtacgtgga cggcgtggag gtgcacaacg ccaagaccaa
     1861 gcccagagag gagcagtaca acagcaccta cagggtggtg tccgtgctga ccgtgctgca
     1921 ccaggactgg ctgaacggca aagaatacaa gtgcaaagtc tccaacaagg ccctggctgc
     1981 cccaatcgaa aagacaatca gcaaggccaa gggccagcca cgggagcccc aggtgtacac
     2041 cctgcccccc agccgggagg agatgaccaa gaaccaggtg tccctgacct gtctggtgaa
     2101 gggcttctac cccagcgata tcgccgtgga gtgggagagc aacggccagc ccgagaacaa
     2161 ctacaagacc acccccccag tgctggacag cgacggcagc ttcttcctgt acagcaagct
     2221 gaccgtggac aagtccaggt ggcagcaggg caacgtgttc agctgcagcg tgatgcacga
     2281 ggccctgcac aaccactaca cccagaagtc cctgagcctg agccccggcg gcagcggtct
     2341 taatgatatt ttcgaggctc agaagattga gtggcatgaa caccaccacc atcaccacta
     2401 aggccggcct gtagtcactg gcacaacgcg ttacccggta agccaatcgg gtatacacgg
     2461 tcgtcatact gcagacaggg ttcttctact ttgcaagata gtcttgagta gtaaaataaa
     2521 tagatagaga aaatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2581 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2641 tgaggaaatt gcatcacaac actcaaccct atctcggtct attcttttga tttataaggg
     2701 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2761 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2881 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2941 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3001 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3061 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3121 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     3181 tatcggcata gtataatacg acaaggtgag gaactaaacc atgaccgagt acaagcccac
     3241 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     3301 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     3361 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     3421 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     3481 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     3541 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     3601 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     3661 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacatccg cgccccgcaa
     3721 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     3781 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cacgtgctac gagatttcga
     3841 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3901 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3961 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4021 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4081 tataccgtcg acctctagct agagcttggc gtaatcatgg tcattaccaa tgcttaatca
     4141 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4201 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac
     4261 cgcgagaacc acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg
     4321 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4381 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta
     4441 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4501 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4561 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4621 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4681 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4741 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4801 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4861 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4921 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4981 tcatattctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5041 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt acaaccaatt
     5101 aaccaattct gaacattatc gcgagcccat ttatacctga atatggctca taacacccct
     5161 tgctcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
     5221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5401 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
     5461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5881 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5941 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6001 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     6061 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     6121 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     6181 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     6241 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     6301 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     6361 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     6421 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     6481 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     6541 ttattatcat ttagaataaa ttttgtgtcg cccttaattg tgagcggata acaattacga
     6601 gcttcatgca cagtggcgtt gactcaatat tggccattag ccatattatt cattggttat
     6661 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     6721 tacatttata ttggctcatg tccaatatga ccgccatgtt
//
