LOCUS       sequence-426450-        6916 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 6916)
  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 6916)
  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-HAVCR1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6916
                     /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             2492..2536
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2537..2554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2694..2711)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2700..2811
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2929..2949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2934..3263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3114..3249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3176..3195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3311..3358
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3377..3976
                     /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(3377..3396)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3873..3893
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4106..4227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4143..4162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4197..4216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4280..5071,5072..5140))
                     /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     4903..4922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5141..5232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5274..5293
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5299..5318
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5404..5992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5893..5912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6146..6163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6261..6278
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6262..6278
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6734..6750
                     /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 cctctgtgaa agttggcgga gaagctggac ctagcgtgac
     1021 cctgccttgt cattattctg gcgccgtgac cagcatgtgc tggaatagag gcagctgcag
     1081 cctgttcacc tgtcagaatg gcatcgtgtg gaccaacggc acccacgtga cctacagaaa
     1141 ggacaccaga tacaagctgc tgggcgacct gagcagacgg gatgtgtctc tgacaatcga
     1201 gaacaccgcc gtgtccgata gcggcgtgta ctgctgtaga gtggaacaca gaggctggtt
     1261 caacgacatg aagatcaccg tgtctctgga aatcgtgccg cctaaagtga ccaccacacc
     1321 tatcgtgacc accgtgccta cagtgacaac cgtgcggaca agcacaacag tgcctaccac
     1381 caccacagtg cccacaacca ctgtgccaac caccatgagc atccccacca caactaccgt
     1441 gctgaccaca atgaccgtgt ccacaacaac cagcgtgccc accactacca gcattcctac
     1501 aaccacatcc gtgcctgtga ccacaaccgt gtctaccttc gtgcctccta tgcctctgcc
     1561 tagacagaat cacgagcctg tggccacctc tcctagtagc cctcagcctg ccgaaacaca
     1621 ccctacaaca ctgcagggcg ccatcagaag agagcctaca agcagccctc tgtacagcta
     1681 caccaccgac ggcaatgata ccgtgaccga gtcctctgac ggcctgtgga acaacaatca
     1741 gacccagctg ttcctggaac actccctgct gaccgccaac accacaaaag gtggcgcgcc
     1801 gggcggcgac aagacccaca cctgcccccc ctgcccagct ccagaactgc tgggagggcc
     1861 ttccgtgttc ctgttccccc ccaagcccaa ggacaccctg atgatcagca ggacccccga
     1921 ggtgacctgc gtggtggtgg ccgtgtccca cgaggaccca gaggtgaagt tcaactggta
     1981 cgtggacggc gtggaggtgc acaacgccaa gaccaagccc agagaggagc agtacaacag
     2041 cacctacagg gtggtgtccg tgctgaccgt gctgcaccag gactggctga acggcaaaga
     2101 atacaagtgc aaagtctcca acaaggccct ggctgcccca atcgaaaaga caatcagcaa
     2161 ggccaagggc cagccacggg agccccaggt gtacaccctg ccccccagcc gggaggagat
     2221 gaccaagaac caggtgtccc tgacctgtct ggtgaagggc ttctacccca gcgatatcgc
     2281 cgtggagtgg gagagcaacg gccagcccga gaacaactac aagaccaccc ccccagtgct
     2341 ggacagcgac ggcagcttct tcctgtacag caagctgacc gtggacaagt ccaggtggca
     2401 gcagggcaac gtgttcagct gcagcgtgat gcacgaggcc ctgcacaacc actacaccca
     2461 gaagtccctg agcctgagcc ccggcggcag cggtcttaat gatattttcg aggctcagaa
     2521 gattgagtgg catgaacacc accaccatca ccactaaggc cggcctgtag tcactggcac
     2581 aacgcgttac ccggtaagcc aatcgggtat acacggtcgt catactgcag acagggttct
     2641 tctactttgc aagatagtct tgagtagtaa aataaataga tagagaaaat cagcctcgac
     2701 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2761 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cacaacactc
     2821 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3181 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3241 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3301 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     3361 ggtgaggaac taaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3421 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3481 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3541 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3601 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3661 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3721 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3781 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3841 ggtgcccgcc ttcctggaga catccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3901 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3961 caagcccggt gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     4021 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4081 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4201 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4261 cttggcgtaa tcatggtcat taccaatgct taatcagtga ggcacctatc tcagcgatct
     4321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4381 agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc
     4441 cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4561 cagttaatag tttgcgcaac gttgttgcca tygctacagg catcgtggtg tcacgctcgt
     4621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt
     5161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5221 aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga
     5281 gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa atcccttaac
     5341 gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5401 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5461 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5521 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt tagcccacca
     5581 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5641 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5701 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5761 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5821 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5881 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5941 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6001 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6061 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6121 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaggcgagag
     6181 tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc tttttgcgtt
     6241 tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg ccccctgggc
     6301 ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct tcggcgggtt
     6361 tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg cgcctgtcac
     6421 tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa cgcactttaa
     6481 ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc atctattatt
     6541 tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga aaataaatct
     6601 cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc aacgataata
     6661 caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag aataaatttt
     6721 gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt ggcgttgact
     6781 caatattggc cattagccat attattcatt ggttatatag cataaatcaa tattggctat
     6841 tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca
     6901 atatgaccgc catgtt
//
