LOCUS       sequence-454865-        9034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein partial 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 9034)
  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 9034)
  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-MXRA5(Trunc3)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..9034
                     /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        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6376..6420
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6421..6438
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6578..6595)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6584..6695
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6813..6833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6818..7147
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6998..7133
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7060..7079
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7195..7242
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7261..7860
                     /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(7261..7280)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7757..7777
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7990..8111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8027..8046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8081..8100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8164..8955,8956..9024))
                     /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     8787..8806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9025..9034,1..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 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgcccc tcagattctg
     2761 acaaagtctc ctcagaccgt gtccgtgacc gccgagacag atacagtgtt tccttgcgag
     2821 gccaccggca agcctaagcc ttttgtgacc tggaccaaag tgtccacagg cgccctgatg
     2881 acccctaaca ccagaatcca gagattcgag gtgctgaaga acggcaccct ggtcatcaga
     2941 aaggtgcagg tccaggacag aggccagtac atgtgtaccg cctctaacct gcacggcctg
     3001 gacagaatgg tggtgctgct gtctgtgaca gtgcagcagc cacagatcct ggccagccac
     3061 tatcaggatg tgaccgtgta cctgggcgac acaatcgcca tggaatgtct ggccaagggc
     3121 acccctgctc ctcagatcag ctggatcttc cccgatcgga gagtgtggca gacagtgtct
     3181 cctgtggaag gcagaatcac cctgcacgag aacagaaccc tgagcatcaa agaggccagc
     3241 ttcagcgacc ggggcgtgta caaatgtgtg gcctctaatg ccgctggcgc cgattctctg
     3301 gccatcagac ttcatgttgc cgctctgcct ccagtgatcc accaagagaa gctggaaaac
     3361 atcagcctgc cacctggcct gtccatccac atccactgta cagccaaagc cgctcctctg
     3421 ccttctgtta gatgggtgct cggcgacggc acacagatca gaccatctca gtttctgcac
     3481 ggcaacctgt tcgtgttccc caatggcacc ctgtacatcc ggaatctggc ccctaaggac
     3541 agcggcagat atgagtgtgt ggccgccaat ctcgtgggca gcgctagaag aactgtgcag
     3601 ctgaatgtgc agagagccgc cgctaacgcc aggattacag gcacaagccc cagacggaca
     3661 gacgtccgat atggcggcac actgaaactg gactgtagcg cctctggcga cccttggcct
     3721 agaatccttt ggagactgcc cagcaagcgg atgatcgacg ccctgttcag cttcgacagc
     3781 cggatcaaag tgttcgccaa cggaacactg gtggtcaaga gcgtgacaga caaggacgcc
     3841 ggcgattatc tgtgcgtggc cagaaacaaa gtgggcgacg actacgtggt gctgaaagtg
     3901 gacgttgtga tgaagcccgc caagatcgag cacaaagaag agaacgacca caaggtgttc
     3961 tacggcggcg atctgaaggt ggactgtgtg gccactggac tgcccaatcc tgagatctct
     4021 tggagcctgc ctgatggcag cctggtcaac agcttcatgc agtccgatga cagcggcggc
     4081 aggaccaaga gatacgtggt gttcaacaac gggaccctgt acttcaacga agtgggcatg
     4141 agagaagagg gcgactacac ctgtttcgcc gagaatcaag tgggcaaaga cgagatgaga
     4201 gtccgcgtga aggttgtgac agcccctgcc accatccgga acaagacata tctggccgtt
     4261 caggtgccat acggcgacgt ggtcacagtg gcttgtgaag ccaaaggcga gcccatgcct
     4321 aaagtgacct ggctgagccc caccaacaaa gtgatcccta ccagcagcga gaagtaccag
     4381 atctaccagg acggaaccct gctgatccag aaggcccaga gaagcgactc cggcaactac
     4441 acatgcctcg tgcgaaattc tgccggcgag gacagaaaga ccgtgtggat tcatgtgaac
     4501 gtgcagcctc caaagatcaa cggcaacccc aatcctatca ccaccgtgcg ggaaattgct
     4561 gctggcggca gcagaaagct gatcgattgc aaggccgagg gcatccccac acctagagtt
     4621 ctgtgggctt ttcctgaggg cgttgtgctg cctgctcctt actacggcaa tcggatcacc
     4681 gtgcatggca acggaagcct ggacatcaga tccctgagaa agtccgacag cgtgcagctc
     4741 gtgtgcatgg ctagaaatga aggcggcgag gccagactga tcctgcagct tacagtgctg
     4801 gaacccatgg aaaagcctat ctttcacgac cccatctccg agaagatcac cgccatggcc
     4861 ggacacacaa tcagcctgaa ttgctctgcc gccggaacac ctacaccttc tcttgtctgg
     4921 gtgctgccca acggcacaga tctgcagtct ggacagcagc tgcagcggtt ctatcacaag
     4981 gccgacggaa tgctgcacat cagcggactg agtagtgtgg acgcaggcgc ttacagatgc
     5041 gtggcacgaa atgccgcagg acacaccgaa agactggtgt ccctgaaagt cggactgaag
     5101 cccgaggcca acaagcagta ccacaacctg gtgtccatca tcaacggcga gacactgaag
     5161 ctgccctgta cacctcctgg tgccggccag ggaagatttt cctggacact gcctaacggc
     5221 atgcacctgg aaggacctca gacactggga agagtgtccc tgctggacaa tggaaccctg
     5281 acagtgcgcg aggcctccgt gtttgataga ggcacctacg tgtgccggat ggaaacagag
     5341 tacggcccca gcgtgaccag cattcctgtg atcgtgatcg cctatcctcc acggatcacc
     5401 agcgagccca cacctgtgat ctacaccaga cctggcaaca ccgtgaagct caactgtatg
     5461 gccatgggca tccctaaggc cgacatcaca tgggagctgc ccgataagtc tcacctgaaa
     5521 gctggcgtgc aggccaggct gtacggcaac agatttctgc atcctcaagg cagcctgacc
     5581 atccagcacg ccacacagag agatgccggc ttctacaagt gcatggccaa gaacatcctg
     5641 ggcagcgaca gcaagaccac ctacatccat gtgtttggcg cgccgggcgg cgacaagacc
     5701 cacacctgcc ccccctgccc agctccagaa ctgctgggag ggccttccgt gttcctgttc
     5761 ccccccaagc ccaaggacac cctgatgatc agcaggaccc ccgaggtgac ctgcgtggtg
     5821 gtggccgtgt cccacgagga cccagaggtg aagttcaact ggtacgtgga cggcgtggag
     5881 gtgcacaacg ccaagaccaa gcccagagag gagcagtaca acagcaccta cagggtggtg
     5941 tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagaatacaa gtgcaaagtc
     6001 tccaacaagg ccctggctgc cccaatcgaa aagacaatca gcaaggccaa gggccagcca
     6061 cgggagcccc aggtgtacac cctgcccccc agccgggagg agatgaccaa gaaccaggtg
     6121 tccctgacct gtctggtgaa gggcttctac cccagcgata tcgccgtgga gtgggagagc
     6181 aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag cgacggcagc
     6241 ttcttcctgt acagcaagct gaccgtggac aagtccaggt ggcagcaggg caacgtgttc
     6301 agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg
     6361 agccccggcg gcagcggtct taatgatatt ttcgaggctc agaagattga gtggcatgaa
     6421 caccaccacc atcaccacta aggccggcct gtagtcactg gcacaacgcg ttacccggta
     6481 agccaatcgg gtatacacgg tcgtcatact gcagacaggg ttcttctact ttgcaagata
     6541 gtcttgagta gtaaaataaa tagatagaga aaatcagcct cgactgtgcc ttctagttgc
     6601 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     6661 actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct atctcggtct
     6721 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6781 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     6841 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6901 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6961 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7021 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7081 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     7141 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     7201 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     7261 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     7321 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     7381 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     7441 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     7501 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     7561 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7621 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     7681 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7741 gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7801 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     7861 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     7921 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     7981 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     8041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     8101 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     8161 tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8221 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8281 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
     8341 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8461 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8581 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8821 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9001 gacacggaaa tgttgaatac tcatattctt cctt
//
