LOCUS       sequence-302829-        7411 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 7411)
  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 7411)
  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-LILRB5-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7411
                     /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             4753..4797
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4798..4815
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4955..4972)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4961..5072
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5190..5210)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5195..5524
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5375..5510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5437..5456
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5572..5619
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5638..6237
                     /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(5638..5657)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6134..6154
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6367..6488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6404..6423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6458..6477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6541..7332,7333..7401))
                     /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     7164..7183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7402..7411,1..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 cggccgccgg aacactgcct
     2761 aaacctacac tttgggccga gcctgcctct gtgattgcca gaggcaaacc cgtgacactg
     2821 tggtgtcagg gccctctgga aaccgaagag taccggctgg acaaagaggg actgccctgg
     2881 gccagaaaga gacagaatcc tctggaacct ggcgccaagg ccaagtttca catccccagc
     2941 acagtgtacg acagcgccgg cagataccgg tgctattatg agacaccagc cggatggtcc
     3001 gagcctagcg atcctcttga actggtggcc acaggcttct acgctgagcc tacactgctt
     3061 gccctgcctt ctcctgttgt ggcctctggc ggaaatgtga ccctgcagtg tgataccctg
     3121 gacggcctgc tgacctttgt gctggtggaa gaggaacaga agctgcccag aacactgtac
     3181 agccagaaac tgcctaaggg cccctctcag gctctgtttc ctgtgggacc tgtgacacct
     3241 agctgcagat ggcggttccg gtgttactac tactaccgga agaaccctca agtgtggtct
     3301 aaccccagcg acctgctgga aattctggtg cctggcgtgt ccagaaagcc ctctttgctg
     3361 attcctcagg gcagcgttgt ggccagaggc ggatctctga cactccagtg tagaagcgac
     3421 gtgggctacg acatcttcgt gctgtacaaa gaaggcgagc acgacctggt gcaaggctct
     3481 ggacaacaac ctcaggctgg cctgtctcag gccaacttta cactgggccc tgtgtccaga
     3541 tctcacggcg gccagtatag atgctacggc gcccataatc tgagccccag gtggtccgct
     3601 cctagcgacc cactggatat tctgatcgcc ggactgatcc ctgacatccc tgctctgtct
     3661 gtgcagcctg gacctaaagt ggccagcggc gagaatgtca cactgctgtg tcagagctgg
     3721 caccagatcg acacattctt tctgacaaaa gagggcgctg cccatcctcc actgtgcctg
     3781 aagtctaagt accagagcta ccggcaccag gccgagttta gcatgtctcc tgtgacatct
     3841 gcccaaggcg gcacctacag atgctactcc gccatcagaa gctaccccta cctgctgagc
     3901 agccccagct atcctcaaga gctggtggtt tctggaccta gcggcgatcc ttctctgtct
     3961 cctaccggct ctacacccac acctggacct gaagatcagc ctctgacacc tactggactg
     4021 gaccctcaat ctggactggg cagacatctc ggaggcgcgc cgggcggcga caagacccac
     4081 acctgccccc cctgcccagc tccagaactg ctgggagggc cttccgtgtt cctgttcccc
     4141 cccaagccca aggacaccct gatgatcagc aggacccccg aggtgacctg cgtggtggtg
     4201 gccgtgtccc acgaggaccc agaggtgaag ttcaactggt acgtggacgg cgtggaggtg
     4261 cacaacgcca agaccaagcc cagagaggag cagtacaaca gcacctacag ggtggtgtcc
     4321 gtgctgaccg tgctgcacca ggactggctg aacggcaaag aatacaagtg caaagtctcc
     4381 aacaaggccc tggctgcccc aatcgaaaag acaatcagca aggccaaggg ccagccacgg
     4441 gagccccagg tgtacaccct gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc
     4501 ctgacctgtc tggtgaaggg cttctacccc agcgatatcg ccgtggagtg ggagagcaac
     4561 ggccagcccg agaacaacta caagaccacc cccccagtgc tggacagcga cggcagcttc
     4621 ttcctgtaca gcaagctgac cgtggacaag tccaggtggc agcagggcaa cgtgttcagc
     4681 tgcagcgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgagc
     4741 cccggcggca gcggtcttaa tgatattttc gaggctcaga agattgagtg gcatgaacac
     4801 caccaccatc accactaagg ccggcctgta gtcactggca caacgcgtta cccggtaagc
     4861 caatcgggta tacacggtcg tcatactgca gacagggttc ttctactttg caagatagtc
     4921 ttgagtagta aaataaatag atagagaaaa tcagcctcga ctgtgccttc tagttgccag
     4981 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     5041 gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt
     5101 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5161 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5221 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5281 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5341 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5401 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5461 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5521 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     5581 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     5641 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5701 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5761 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5821 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5881 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5941 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6001 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6061 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6121 acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6181 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac
     6241 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6301 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6361 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6421 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6481 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     6541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6661 cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa
     6721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6841 cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7381 acggaaatgt tgaatactca tattcttcct t
//
