LOCUS       sequence-412941-        9157 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 9157)
  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 9157)
  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-ISLR2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..9157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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        3105..3484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3485..3688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3639..3659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3685..3709
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3919..3937
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3919..3936
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     3950..3969
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6499..6543
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6544..6561
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6701..6718)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6707..6818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6936..6956)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6941..7270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7121..7256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        7318..7365
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7384..7983
                     /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(7384..7403)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7880..7900
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8113..8234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8150..8169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8204..8223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8287..9078,9079..9147))
                     /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     8910..8929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9148..9157,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 gctgagagat cccctcataa tttccccaaa gcgtaaccat gtgtgaataa attttgagct
     1681 agtagggttg cagccacgag taagtcttcc cttgttattg tgtagccaga atgccgcaaa
     1741 acttccatgc ctaagcgaac tgttgagagt acgtttcgat ttctgactgt gttagcctgg
     1801 aagtgcttgt cccaaccttg tttctgagca tgaacgcccg caagccaaca tgttagttga
     1861 agcatcaggg cgattagcag catgatatca aaacgctctg agctgctcgt tcggctatgg
     1921 cgtaggccta gtccgtaggc aggacttttc aagtctcgga aggtttcttc aatctgcatt
     1981 cgcttcgaat agatattaac aagttgtttg ggtgttcgaa tttcaacagg taagttagtt
     2041 gctagaaccc atggctcctt tgccgacgct gagtagattt taggtgacgg gtggtgacaa
     2101 tgagtccgtg tcgagcgctg attttttcgg cctttagagc gagatttata caatagaatt
     2161 tggcatgaga ttggattgct tttagtcagc ctcttatagc ctaaagtctt tgagtgacta
     2221 gatgacatat catgtaagtt gctgataggt ttccagtttt ccgctcctag gtctgcatat
     2281 tgtacttttc ctcttactcg acttaaccag taccaaccca gcttctcaac ggatttatac
     2341 catggcactt taaagccagc atcactgaca atgagcggtg tggtgttact cggtagaatg
     2401 ctcgcaaggt cggctagaaa ttggtcatga gctttctttg aacattgctc tgaaagcggg
     2461 aacgctttct cataaagagt aacagaacga ccgtgtagtg cgactgaagc tcgcaatacc
     2521 ataagtcgtt tttgctcacg aatatcagac cagtcaacaa gtacaatggg catcgtattg
     2581 cccgaacaga taaagctagc atgccaacgg tatacagcga gtcgctcttt gtggaggtga
     2641 cgattaccta acaatcggtc gattcgtttg atgttatgtt ttgttctcgc tttggttggc
     2701 aggttacggc caagttcggt aagagtgaga gttttacagt caagtaatgc gtggcaagcc
     2761 aacgttaagc tgttgagtcg ttttaagtgt aattcggggc agaattggta aagagagtcg
     2821 tgtaaaatat cgagttcgca catcttgttg tctgattatt gatttttcgc gaaaccattt
     2881 gatcatatga caagatgtgt atccacctta acttaatgat ttttaccaaa atcattaggg
     2941 gattcatcag tgcacagtgg cgttgactca atattggcca ttagccatat tattcattgg
     3001 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     3061 tatgtacatt tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac
     3121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     3181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     3241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     3301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     3541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     3721 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
     3781 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
     3841 agtctatagg cccaccccct tggcttcgtt agaacgcggc tacaattaat acataacctt
     3901 atgtatcata cacatacgat ttaggtgaca ctatagaata acatccactt tgcctttctc
     3961 tccacaggtg tccactccca ggtccaactg cacctcggtt ctatcgaaaa cgcgcctgct
     4021 agcgccacca tgaagaccat cattgccctg agctacatct tctgcctggt gttcgctgcg
     4081 gccgcctctt gtcctgaacc ttgtgcctgc gtggacaaat acgcccacca gttcgccgac
     4141 tgcgcctaca aagaactgag agaggtgcca gagggcctgc ctgccaatgt gacaacactg
     4201 agcctgagcg ccaacaagat caccgtgctg agaagaggcg ccttcgccga tgtgacccaa
     4261 gtgacatctc tgtggctggc ccacaacgaa gtgcggacag ttgaacctgg tgctctggcc
     4321 gttctgagcc agctgaagaa cctggacctg agccacaact tcatcagcag cttcccttgg
     4381 agcgacctgc ggaatctgtc tgctctgcag ctgctgaaga tgaaccacaa ccggctggga
     4441 agcctgccta gagatgctct gggagcactg cccgatctgc ggagcctgag aatcaacaac
     4501 aacagactgc ggacactggc cccaggcacc tttgatgctc tgagcgctct gtctcacctc
     4561 cagctgtatc acaacccctt ccactgtggc tgtggcctcg tttggcttca agcctgggcc
     4621 gccagtacca gagtgtctct gcctgagcca gactctatcg cctgtgcctc tccaccagct
     4681 ctgcagggcg ttccagtgta cagacttcct gctctgcctt gcgctcctcc tagcgttcac
     4741 ctgtctgccg aacctccact ggaagcccct ggaacacctc tgagagccgg actggctttc
     4801 gtgctgcact gtatcgccga cggacacccc acacctagac tgcagtggca gctgcaaatc
     4861 cctggcggaa cagtggtgct ggaacctcct gttctgagcg gcgaagatga tggcgtggga
     4921 gccgaagaag gcgaaggcga aggcgacggg gatctgctga cacagacaca ggctcagacc
     4981 cctacaccag ctccagcttg gcctgctcct cctgccacac caagatttct ggccctggcc
     5041 aatggcagcc tgctggttcc tctgctgtct gccaaagagg ccggcgtcta cacatgcaga
     5101 gcccacaatg agctgggcgc caatagcacc tctatcagag tggccgtggc cgccacagga
     5161 cctccaaaac atgctcctgg tgctggcgga gagcctgatg gacaagcccc tacaagcgag
     5221 agaaagagca ccgccaaagg cagaggcaac agcgtgctgc cttctaagcc tgagggcaag
     5281 atcaaaggcc agggactcgc caaggtgtcc atcctgggag aaaccgagac tgagcccgaa
     5341 gaggatacca gcgagggcga agaggccgag gatcagattc tggctgatcc cgccgaggaa
     5401 cagagatgcg gaaacggcga tcccagcaga tacgtgtcca accacgcctt caatcagagc
     5461 gccgagctga agcctcacgt gttcgagctg ggagtgatcg ccctggatgt ggctgaaagg
     5521 gaagccagag tgcagctgac accactggct gctagatggg gacctggacc aggcggtgca
     5581 ggcggagcac ctagacctgg aagaaggcct ctgagactgc tgtatctgtg tccagctggt
     5641 ggcggagctg ctgtgcaatg gtccagagtt gaggaaggcg tgaacgccta ctggttcaga
     5701 ggactgaggc ccggcaccaa ctactctgtg tgtcttgctc tggctggcga ggcctgtcat
     5761 gtgcaagtgg tgttcagcac caagaaagag ctgccctctg gcgcgccggg cggcgacaag
     5821 acccacacct gccccccctg cccagctcca gaactgctgg gagggccttc cgtgttcctg
     5881 ttccccccca agcccaagga caccctgatg atcagcagga cccccgaggt gacctgcgtg
     5941 gtggtggccg tgtcccacga ggacccagag gtgaagttca actggtacgt ggacggcgtg
     6001 gaggtgcaca acgccaagac caagcccaga gaggagcagt acaacagcac ctacagggtg
     6061 gtgtccgtgc tgaccgtgct gcaccaggac tggctgaacg gcaaagaata caagtgcaaa
     6121 gtctccaaca aggccctggc tgccccaatc gaaaagacaa tcagcaaggc caagggccag
     6181 ccacgggagc cccaggtgta caccctgccc cccagccggg aggagatgac caagaaccag
     6241 gtgtccctga cctgtctggt gaagggcttc taccccagcg atatcgccgt ggagtgggag
     6301 agcaacggcc agcccgagaa caactacaag accacccccc cagtgctgga cagcgacggc
     6361 agcttcttcc tgtacagcaa gctgaccgtg gacaagtcca ggtggcagca gggcaacgtg
     6421 ttcagctgca gcgtgatgca cgaggccctg cacaaccact acacccagaa gtccctgagc
     6481 ctgagccccg gcggcagcgg tcttaatgat attttcgagg ctcagaagat tgagtggcat
     6541 gaacaccacc accatcacca ctaaggccgg cctgtagtca ctggcacaac gcgttacccg
     6601 gtaagccaat cgggtataca cggtcgtcat actgcagaca gggttcttct actttgcaag
     6661 atagtcttga gtagtaaaat aaatagatag agaaaatcag cctcgactgt gccttctagt
     6721 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6781 cccactgtcc tttcctaata aaatgaggaa attgcatcac aacactcaac cctatctcgg
     6841 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6901 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     6961 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     7021 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     7081 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     7141 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     7201 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     7261 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     7321 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     7381 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     7441 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7501 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7561 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     7621 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     7681 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     7741 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     7801 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     7861 ctggagacat ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     7921 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     7981 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     8041 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     8101 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     8161 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8221 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     8281 tggtcattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8401 tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc
     8461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8581 gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9121 ggcgacacgg aaatgttgaa tactcatatt cttcctt
//
