LOCUS       sequence-371820-        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-TMIGD2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(11..102)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     144..163
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     169..188
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      274..862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     763..782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1016..1033
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1131..1148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1132..1148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1604..1620
                     /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        1787..2166
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2167..2370
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2321..2341
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2367..2391
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2601..2619
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2601..2618
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2632..2651
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3852..3896
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3897..3914
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4054..4071)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4060..4171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4289..4309)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4294..4623
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4474..4609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4671..4718
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4737..5336
                     /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(4737..4756)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5233..5253
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5466..5587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5557..5576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5640..6431,6432..6490,1..10))
                     /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     6263..6282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
       61 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
      121 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
      181 accccttgct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
      241 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      301 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      361 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      421 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
      481 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      541 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      601 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      661 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      721 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      781 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      841 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      901 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      961 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     1021 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     1081 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     1141 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     1201 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     1261 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     1321 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     1381 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     1441 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     1501 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     1561 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     1621 ttacgagctt catgcacagt ggcgttgact caatattggc cattagccat attattcatt
     1681 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     1741 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     1801 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     1861 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1921 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     1981 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     2041 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     2101 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     2161 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     2221 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     2281 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     2341 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     2401 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccgcggc
     2461 cgggaacggt gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat
     2521 agagtctata ggcccacccc cttggcttcg ttagaacgcg gctacaatta atacataacc
     2581 ttatgtatca tacacatacg atttaggtga cactatagaa taacatccac tttgcctttc
     2641 tctccacagg tgtccactcc caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg
     2701 ctagcgccac catgaagacc atcattgccc tgagctacat cttctgcctg gtgttcgctg
     2761 cggccgcctt gtctgttcag cagggaccta atctgctgca agtgcggcag ggatctcagg
     2821 ccacactcgt gtgtcaagtg gatcaggcta cagcctggga gagactgaga gtgaagtgga
     2881 caaaggacgg ggccatcctg tgccagcctt acatcacaaa tggcagcctg agcctgggcg
     2941 tgtgtggacc tcaaggcaga ctttcttggc aggccccttc tcacctgaca ctgcagctgg
     3001 accctgtgtc tctgaatcac agcggcgcct atgtgtgttg ggccgctgtg gaaattcctg
     3061 agctggaaga ggccgagggc aacatcacca gactgttcgt ggaccccgac gatcccacac
     3121 agaaccggaa cagaattgcc agctttcccg gtggcgcgcc gggcggcgac aagacccaca
     3181 cctgcccccc ctgcccagct ccagaactgc tgggagggcc ttccgtgttc ctgttccccc
     3241 ccaagcccaa ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg
     3301 ccgtgtccca cgaggaccca gaggtgaagt tcaactggta cgtggacggc gtggaggtgc
     3361 acaacgccaa gaccaagccc agagaggagc agtacaacag cacctacagg gtggtgtccg
     3421 tgctgaccgt gctgcaccag gactggctga acggcaaaga atacaagtgc aaagtctcca
     3481 acaaggccct ggctgcccca atcgaaaaga caatcagcaa ggccaagggc cagccacggg
     3541 agccccaggt gtacaccctg ccccccagcc gggaggagat gaccaagaac caggtgtccc
     3601 tgacctgtct ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagagcaacg
     3661 gccagcccga gaacaactac aagaccaccc ccccagtgct ggacagcgac ggcagcttct
     3721 tcctgtacag caagctgacc gtggacaagt ccaggtggca gcagggcaac gtgttcagct
     3781 gcagcgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg agcctgagcc
     3841 ccggcggcag cggtcttaat gatattttcg aggctcagaa gattgagtgg catgaacacc
     3901 accaccatca ccactaaggc cggcctgtag tcactggcac aacgcgttac ccggtaagcc
     3961 aatcgggtat acacggtcgt catactgcag acagggttct tctactttgc aagatagtct
     4021 tgagtagtaa aataaataga tagagaaaat cagcctcgac tgtgccttct agttgccagc
     4081 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4141 tcctttccta ataaaatgag gaaattgcat cacaacactc aaccctatct cggtctattc
     4201 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4261 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4321 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4441 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4501 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4561 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4621 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     4681 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga
     4741 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4801 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4861 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4921 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4981 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     5041 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5101 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5161 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5221 catccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5281 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg
     5341 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5401 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5461 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5521 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5581 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5761 agcgctgcga tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac
     5821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5941 gttgttgcca tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6481 cggaaatgtt
//
