LOCUS       sequence-368528-        6589 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 6589)
  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 6589)
  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-EDAR-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..120
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(139..186)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(234..563)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      248..383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(302..321)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     548..568
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    686..797
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     786..803
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(943..960)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(961..1005)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2305..2324)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2337..2355)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2338..2355)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2565..2589)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2586..2789)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2615..2635)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2790..3169
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3336..3352
                     /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)."
     primer_bind     complement(3808..3825)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3808..3824)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3923..3940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4094..4682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4174..4193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4768..4787)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4793..4812)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4854..4945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4946..5014,5015..5806)
                     /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     complement(5164..5183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5859..5980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5870..5889)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5924..5943
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6110..6589,1..120))
                     /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(6193..6213)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
       61 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
      121 ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt
      181 gtcaacacgt gctgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa
      241 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg
      301 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
      361 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      421 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      481 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      541 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
      601 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      661 agaccgagat agggttgagt gttgtgatgc aatttcctca ttttattagg aaaggacagt
      721 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
      781 gcaactagaa ggcacagtcg aggctgattt tctctatcta tttattttac tactcaagac
      841 tatcttgcaa agtagaagaa ccctgtctgc agtatgacga ccgtgtatac ccgattggct
      901 taccgggtaa cgcgttgtgc cagtgactac aggccggcct tagtggtgat ggtggtggtg
      961 ttcatgccac tcaatcttct gagcctcgaa aatatcatta agaccgctgc cgccggggct
     1021 caggctcagg gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct
     1081 gaacacgttg ccctgctgcc acctggactt gtccacggtc agcttgctgt acaggaagaa
     1141 gctgccgtcg ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt
     1201 gctctcccac tccacggcga tatcgctggg gtagaagccc ttcaccagac aggtcaggga
     1261 cacctggttc ttggtcatct cctcccggct ggggggcagg gtgtacacct ggggctcccg
     1321 tggctggccc ttggccttgc tgattgtctt ttcgattggg gcagccaggg ccttgttgga
     1381 gactttgcac ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga
     1441 caccaccctg taggtgctgt tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac
     1501 ctccacgccg tccacgtacc agttgaactt cacctctggg tcctcgtggg acacggccac
     1561 caccacgcag gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg gcttgggggg
     1621 gaacaggaac acggaaggcc ctcccagcag ttctggagct gggcaggggg ggcaggtgtg
     1681 ggtcttgtcg ccgcccggcg cgccagcggt ggccagatgt ccctgtccag acagctcttt
     1741 gtgggcgtgc tgaaaagggc tcagtgtgct gcttccgctt gtgccaggaa aattggcgct
     1801 ggcgccgctt gtggctccca cacattcttt ggtgttagga ggggccagca gacaagagta
     1861 gcagaccatg ccgtagatgt tccggggtct gttttccagc atgtagtagc caggcaggca
     1921 aggtccacac tcggcatcat tttccatgtc gccaggggtc agcactgtgg ctctaaagaa
     1981 gccctcgcag tccttgtgcc gtctgcagat ctggtagccg cctttgctga acttctcggc
     2041 aggacaaggc acacagccgt aatcctcatc cttggtgccg tagccacagc tcaggtaagg
     2101 ttcctcgcca ggtccacaag gaggacactc ttggcacagg ccggtggtct ggttgtagta
     2161 ctcgttctcg ccgcaattgc tatactcggc ggccgcagcg aacaccaggc agaagatgta
     2221 gctcagggca atgatggtct tcatggtggc gctagcaggc gcgttttcga tagaaccgag
     2281 gtgcagttgg acctgggagt ggacacctgt ggagagaaag gcaaagtgga tgttattcta
     2341 tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt
     2401 tctaacgaag ccaagggggt gggcctatag actctatagg cggtacttac gtcactcttg
     2461 gcacggggaa tccgcgttcc aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg
     2521 tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact
     2581 aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     2641 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     2701 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     2761 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     2821 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     2881 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     2941 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     3001 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3061 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3121 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgcca acatggcggt
     3181 catattggac atgagccaat ataaatgtac atattatgat atagatacaa cgtatgcaat
     3241 ggccaatagc caatattgat ttatgctata taaccaatga ataatatggc taatggccaa
     3301 tattgagtca acgccactgt gcatgaagct cgtaattgtt atccgctcac aattaagggc
     3361 gacacaaaat ttattctaaa tgataataaa tactgataac atcttatagt ttgtattata
     3421 ttttgtatta tcgttgacat gtataatttt gatatcaaaa actgattttc cctttattat
     3481 tttcgagatt tattttctta attctcttta acaaactaga aatattgtat atacaaaaaa
     3541 tcataaataa tagatgaata gtttaattat aggtgttcat caatcgaaaa agcaacgtat
     3601 cttatttaaa gtgcgttgct tttttctcat ttataaggtt aaataattct catatatcaa
     3661 gcaaagtgac aggcgccctt aaatattctg acaaatgctc tttccctaaa ctccccccat
     3721 aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca
     3781 gaaccgccca gggggcccga gcttaagact ggccgtcgtt ttacaacaca gaaagagttt
     3841 gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct tagtttgatg cctggcagtt
     3901 ccctactctc gccttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3961 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4021 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4501 tgaagtggtg ggctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4561 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4621 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4681 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa
     4741 ctcacgttaa gggattttgg tcatgagcaa ggggtgttat gagccatatt caggtataaa
     4801 tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt
     4861 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4921 tcaataatat tgaaaaagga agaatatgag tattcaacat ttccgtgtcg cccttattcc
     4981 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5041 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5101 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5161 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5221 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5281 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5341 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5401 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5461 aaacgacgag cgtgacacca cgatgcctgt agcgatggca acaacgttgc gcaaactatt
     5521 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5581 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5641 atccggagcc ggtgagcgtg gttctcgcgg tatcatcgca gcgctggggc cagatggtaa
     5701 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5761 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaatgac catgattacg
     5821 ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg
     5881 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5941 tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca
     6001 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     6061 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg
     6121 gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg
     6181 tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg
     6241 gcaccccggc gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct
     6301 tgccctggtg gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc tccttgggcc
     6361 ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact
     6421 cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg
     6481 tccagaccgc caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga
     6541 cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc gatgtggcg
//
