LOCUS       sequence-401536-        7150 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 7150)
  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 7150)
  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-CD5-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7150
                     /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             2726..2770
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2771..2788
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2928..2945)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2934..3045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3163..3183)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3168..3497
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3348..3483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3410..3429
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3545..3592
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3611..4210
                     /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(3611..3630)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4107..4127
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4340..4461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4377..4396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4431..4450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4514..5305,5306..5374))
                     /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     5137..5156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5375..5466)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5508..5527
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5533..5552
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5638..6226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6127..6146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6380..6397
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6495..6512
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6496..6512
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6968..6984
                     /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 ccagactttc ttggtacgac cccgatttcc aggccagact
     1021 gacccggtcc aatagcaagt gtcagggcca gctggaagtg tacctgaagg acggatggca
     1081 catggtctgc agccagagct ggggcagatc tagcaagcag tgggaagatc ctagccaggc
     1141 cagcaaagtg tgccagagac tgaattgcgg cgtgcccctg tctctgggcc ctttcctggt
     1201 cacatacacc cctcagagca gcatcatctg ctacggacag ctgggcagct tcagcaactg
     1261 cagccacagc agaaacgaca tgtgccacag cctgggcctg acctgcctgg aacctcagaa
     1321 aaccacacct cctaccacca gacctccacc aaccaccaca cctgagccta ccgctcctcc
     1381 tagactgcag ctggttgctc aatctggcgg acagcattgt gccggcgtgg tggaattcta
     1441 ctctggctct ctcggcggca ccatcagcta tgaggcccag gataagaccc aggacctgga
     1501 aaacttcctg tgcaacaacc tgcagtgcgg cagcttcctg aaacatctgc ccgagacaga
     1561 agccggcaga gcacaagatc ctggcgagcc tagagaacac cagcctctgc ctatccagtg
     1621 gaagattcag aacagcagct gcaccagcct ggaacactgc ttcagaaaga tcaagcccca
     1681 gaaaagcggc agagtgctgg ctctgctgtg cagcggattt cagcctaagg tgcagagcag
     1741 actcgtcggc ggaagcagca tctgtgaagg cacagtggaa gttcggcagg gcgctcaatg
     1801 ggccgctctg tgtgattcta gcagcgccag atccagcctg agatgggaag aagtgtgccg
     1861 cgagcagcag tgtggcagcg tgaacagcta cagagtcctg gatgccggcg atcctacaag
     1921 cagaggcctg ttctgccctc accagaaact gagccagtgc cacgagctgt gggagagaaa
     1981 cagctactgc aagaaagtgt tcgtgacctg tcaggacccc aatcctggcg cgccgggcgg
     2041 cgacaagacc cacacctgcc ccccctgccc agctccagaa ctgctgggag ggccttccgt
     2101 gttcctgttc ccccccaagc ccaaggacac cctgatgatc agcaggaccc ccgaggtgac
     2161 ctgcgtggtg gtggccgtgt cccacgagga cccagaggtg aagttcaact ggtacgtgga
     2221 cggcgtggag gtgcacaacg ccaagaccaa gcccagagag gagcagtaca acagcaccta
     2281 cagggtggtg tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagaatacaa
     2341 gtgcaaagtc tccaacaagg ccctggctgc cccaatcgaa aagacaatca gcaaggccaa
     2401 gggccagcca cgggagcccc aggtgtacac cctgcccccc agccgggagg agatgaccaa
     2461 gaaccaggtg tccctgacct gtctggtgaa gggcttctac cccagcgata tcgccgtgga
     2521 gtgggagagc aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag
     2581 cgacggcagc ttcttcctgt acagcaagct gaccgtggac aagtccaggt ggcagcaggg
     2641 caacgtgttc agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc
     2701 cctgagcctg agccccggcg gcagcggtct taatgatatt ttcgaggctc agaagattga
     2761 gtggcatgaa caccaccacc atcaccacta aggccggcct gtagtcactg gcacaacgcg
     2821 ttacccggta agccaatcgg gtatacacgg tcgtcatact gcagacaggg ttcttctact
     2881 ttgcaagata gtcttgagta gtaaaataaa tagatagaga aaatcagcct cgactgtgcc
     2941 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3001 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct
     3061 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3121 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3241 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3301 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3361 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3421 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3481 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     3541 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3601 gaactaaacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     3661 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     3721 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     3781 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     3841 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     3901 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     3961 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4021 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4081 cgccttcctg gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4141 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4201 cggtgcctga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4261 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4321 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4381 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4441 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4501 gtaatcatgg tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4621 taccatctgg ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt
     4681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4801 atagtttgcg caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5341 gaataagggc gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa
     5401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5461 aacaaatagg ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat
     5521 ttatacctga atatggctca taacacccct tgctcatgac caaaatccct taacgtgagt
     5581 tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa
     5821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6301 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6361 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga
     6421 actgccaggc atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca
     6481 aactctttct gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct
     6541 gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta
     6601 tggggggagt ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct
     6661 tgatatatga gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga
     6721 tacgttgctt tttcgattga tgaacaccta taattaaact attcatctat tatttatgat
     6781 tttttgtata tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa
     6841 taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat
     6901 ataatacaaa ctataagatg ttatcagtat ttattatcat ttagaataaa ttttgtgtcg
     6961 cccttaattg tgagcggata acaattacga gcttcatgca cagtggcgtt gactcaatat
     7021 tggccattag ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca
     7081 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
     7141 ccgccatgtt
//
