LOCUS       sequence-368829-        6475 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 6475)
  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 6475)
  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-CD47-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     423..443
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    656..777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(693..712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     747..766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(830..1621,1622..1690))
                     /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     1453..1472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1691..1782)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1824..1843
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1849..1868
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1954..2542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2443..2462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2696..2713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2811..2828
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2812..2828
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3284..3300
                     /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        3467..3846
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3847..4050
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4001..4021
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4047..4071
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4281..4299
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4281..4298
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4312..4331
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5517..5561
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5562..5579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5719..5736)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5725..5836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5954..5974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5959..6288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6139..6274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6201..6220
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6336..6383
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(6402..6475,1..526)
                     /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(6402..6421)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
       61 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
      121 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
      181 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
      241 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
      301 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
      361 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga catccgcgcc
      421 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
      481 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg tgctacgaga
      541 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
      601 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
      661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      781 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat taccaatgct
      841 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      901 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
      961 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     1021 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1081 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1141 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1201 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1261 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1321 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1381 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1441 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1501 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1561 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1621 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1681 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1741 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
     1801 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
     1861 accccttgct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     1921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2101 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     2161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2701 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     2761 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     2821 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     2881 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     2941 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     3001 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     3061 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     3121 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     3181 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     3241 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     3301 ttacgagctt catgcacagt ggcgttgact caatattggc cattagccat attattcatt
     3361 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     3421 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     3481 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3541 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3601 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3661 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3721 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3781 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     3841 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     3901 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     3961 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     4021 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     4081 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccgcggc
     4141 cgggaacggt gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat
     4201 agagtctata ggcccacccc cttggcttcg ttagaacgcg gctacaatta atacataacc
     4261 ttatgtatca tacacatacg atttaggtga cactatagaa taacatccac tttgcctttc
     4321 tctccacagg tgtccactcc caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg
     4381 ctagcgccac catgaagacc atcattgccc tgagctacat cttctgcctg gtgttcgctg
     4441 cggccgccca actgctgttc aacaagacca agagcgtgga attcaccttc tgcaacgaca
     4501 ccgtggtcat cccttgcttc gtgaccaaca tggaagccca gaacaccacc gaggtgtacg
     4561 tgaagtggaa gttcaagggc agagacatct acaccttcga cggcgccctg aacaagagca
     4621 ccgtgcctac agatttcagc agcgccaaga tcgaggtgtc ccagctgctt aaaggcgacg
     4681 ccagcctgaa gatggataag agcgacgccg tgtctcacac cggcaactac acctgtgaag
     4741 tgaccgagct gaccagagag ggcgagacaa tcatcgagct gaagtacaga gtggtgtcct
     4801 ggttcagccc caacgaaggc gcgccgggcg gcgacaagac ccacacctgc cccccctgcc
     4861 cagctccaga actgctggga gggccttccg tgttcctgtt cccccccaag cccaaggaca
     4921 ccctgatgat cagcaggacc cccgaggtga cctgcgtggt ggtggccgtg tcccacgagg
     4981 acccagaggt gaagttcaac tggtacgtgg acggcgtgga ggtgcacaac gccaagacca
     5041 agcccagaga ggagcagtac aacagcacct acagggtggt gtccgtgctg accgtgctgc
     5101 accaggactg gctgaacggc aaagaataca agtgcaaagt ctccaacaag gccctggctg
     5161 ccccaatcga aaagacaatc agcaaggcca agggccagcc acgggagccc caggtgtaca
     5221 ccctgccccc cagccgggag gagatgacca agaaccaggt gtccctgacc tgtctggtga
     5281 agggcttcta ccccagcgat atcgccgtgg agtgggagag caacggccag cccgagaaca
     5341 actacaagac caccccccca gtgctggaca gcgacggcag cttcttcctg tacagcaagc
     5401 tgaccgtgga caagtccagg tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg
     5461 aggccctgca caaccactac acccagaagt ccctgagcct gagccccggc ggcagcggtc
     5521 ttaatgatat tttcgaggct cagaagattg agtggcatga acaccaccac catcaccact
     5581 aaggccggcc tgtagtcact ggcacaacgc gttacccggt aagccaatcg ggtatacacg
     5641 gtcgtcatac tgcagacagg gttcttctac tttgcaagat agtcttgagt agtaaaataa
     5701 atagatagag aaaatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5761 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5821 atgaggaaat tgcatcacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5881 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5941 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6061 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6121 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6181 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     6241 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     6301 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     6361 atatcggcat agtataatac gacaaggtga ggaactaaac catgaccgag tacaagccca
     6421 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgc
//
