LOCUS       sequence-401040-        7159 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 7159)
  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 7159)
  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-UNC5B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7159
                     /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             2735..2779
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2780..2797
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2937..2954)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2943..3054
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3172..3192)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3177..3506
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3357..3492
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3419..3438
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3554..3601
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3620..4219
                     /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(3620..3639)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4116..4136
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4349..4470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4386..4405)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4440..4459
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4523..5314,5315..5383))
                     /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     5146..5165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5384..5475)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5517..5536
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5542..5561
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5647..6235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6136..6155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6389..6406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6504..6521
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6505..6521
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6977..6993
                     /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 ccggaacaga ttctggatct gaggttctgc ccgacagctt
     1021 ccctagcgct cctgctgaac ccctgcctta cttcctgcaa gagccccagg atgcctacat
     1081 cgtgaagaac aagcccgtgg aactgcggtg cagagccttt cctgccacac agatctactt
     1141 caagtgcaac ggcgagtggg tgtcccagaa cgaccacgtg acacaagaag gactggacga
     1201 ggccacaggc ctgagagtca gagaagtgca gatcgaggtg tcccggcagc aggtcgagga
     1261 actgtttggc ctggaagatt actggtgcca gtgcgtggca tggtctagcg ccggcaccac
     1321 aaaaagcaga cgggcctacg tgcggatcgc ctacctgcgg aagaacttcg accaagagcc
     1381 tctgggcaaa gaggtgcccc tggatcacga agtgctgctg cagtgtagac ctcctgaagg
     1441 cgtgccagtg gccgaagtgg aatggctgaa gaacgaggac gtgatcgacc ctactcagga
     1501 taccaacttc ctgctgacca tcgaccacaa cctgatcatc cggcaggcca gactgagcga
     1561 caccgccaat tatacctgcg tggccaagaa cattgtggcc aagcggagaa gcaccaccgc
     1621 caccgtgatc gtgtatgtga atggcggctg gtctagctgg gccgagtggt ccccttgcag
     1681 caatagatgt ggcagaggct ggcagaagcg gaccagaacc tgtacaaacc ccgcacctct
     1741 taacggcgga gccttctgtg aaggccaggc ctttcagaaa accgcctgca ccaccatctg
     1801 tcccgttgat ggtgcttgga cagagtggtc caagtggtcc gcctgtagca cagaatgcgc
     1861 ccattggcgg agccgcgagt gtatggctcc tccacctcaa aacggcggca gagattgctc
     1921 tggcaccctg ctggacagca agaactgtac cgatggcctg tgcatgcaga acaagaaaac
     1981 cctgagcgac cccaacagcc atctgctgga agccagcgga gatgccgctc tgtatggcgc
     2041 gccgggcggc gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg
     2101 gccttccgtg ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc
     2161 cgaggtgacc tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg
     2221 gtacgtggac ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa
     2281 cagcacctac agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa
     2341 agaatacaag tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag
     2401 caaggccaag ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga
     2461 gatgaccaag aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat
     2521 cgccgtggag tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt
     2581 gctggacagc gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg
     2641 gcagcagggc aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac
     2701 ccagaagtcc ctgagcctga gccccggcgg cagcggtctt aatgatattt tcgaggctca
     2761 gaagattgag tggcatgaac accaccacca tcaccactaa ggccggcctg tagtcactgg
     2821 cacaacgcgt tacccggtaa gccaatcggg tatacacggt cgtcatactg cagacagggt
     2881 tcttctactt tgcaagatag tcttgagtag taaaataaat agatagagaa aatcagcctc
     2941 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3001 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcacaaca
     3061 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3121 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3181 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3241 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3301 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3361 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3421 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3481 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3541 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3601 caaggtgagg aactaaacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     3661 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     3721 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     3781 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     3841 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3901 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3961 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     4021 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4081 cggggtgccc gccttcctgg agacatccgc gccccgcaac ctccccttct acgagcggct
     4141 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4201 ccgcaagccc ggtgcctgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     4261 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4321 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4381 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4441 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4501 gagcttggcg taatcatggt cattaccaat gcttaatcag tgaggcacct atctcagcga
     4561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4621 gggagggctt accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg
     4681 ctccggattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4801 cgccagttaa tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct
     4861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5341 caaaaaaggg aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat
     5401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5461 agaaaaataa acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg
     5521 cgagcccatt tatacctgaa tatggctcat aacacccctt gctcatgacc aaaatccctt
     5581 aacgtgagtt acgcgcgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttagccca
     5821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6241 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6301 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6361 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga
     6421 gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg gcctttttgc
     6481 gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc gggccccctg
     6541 ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg
     6601 gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa gggcgcctgt
     6661 cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag caacgcactt
     6721 taaataagat acgttgcttt ttcgattgat gaacacctat aattaaacta ttcatctatt
     6781 atttatgatt ttttgtatat acaatatttc tagtttgtta aagagaatta agaaaataaa
     6841 tctcgaaaat aataaaggga aaatcagttt ttgatatcaa aattatacat gtcaacgata
     6901 atacaaaata taatacaaac tataagatgt tatcagtatt tattatcatt tagaataaat
     6961 tttgtgtcgc ccttaattgt gagcggataa caattacgag cttcatgcac agtggcgttg
     7021 actcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
     7081 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
     7141 ccaatatgac cgccatgtt
//
