LOCUS       sequence-410016-        7381 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 7381)
  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 7381)
  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-SIGLEC5-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7381
                     /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             2957..3001
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3002..3019
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3159..3176)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3165..3276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3394..3414)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3399..3728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3579..3714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3641..3660
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3776..3823
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3842..4441
                     /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(3842..3861)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4338..4358
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4571..4692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4608..4627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4662..4681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4745..5536,5537..5605))
                     /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     5368..5387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5606..5697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5739..5758
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5764..5783
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5869..6457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6358..6377
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6611..6628
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6726..6743
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6727..6743
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7199..7215
                     /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 ccgaaaagcc tgtgtatgag ctgcaggtcc agaaaagcgt
     1021 gaccgtgcaa gagggcctgt gtgtgctggt gccttgcagc tttagctacc cttggcggag
     1081 ctggtacagc agccctcctc tgtacgtgta ctggttcaga gatggcgaga tcccctacta
     1141 cgccgaggtg gtggccacaa acaaccccga cagaagagtg aagcccgaga cacagggcag
     1201 attcagactg ctgggcgacg tgcagaagaa gaactgcagc ctgagcatcg gcgacgcccg
     1261 gatggaagat accggcagct acttcttcag agtggaacgc ggcagggacg tgaagtacag
     1321 ctaccagcag aacaagctga acctggaagt gacagccctg atcgagaagc ccgacatcca
     1381 ctttctggaa cccctggaaa gcggcagacc caccagactg tcttgtagcc tgcctggcag
     1441 ctgtgaagct ggccctccac tgacctttag ctggaccggc aatgccctgt ctcctctgga
     1501 ccctgagaca accagaagca gcgagctgac cctgacacct agacctgagg accacggcac
     1561 caatctgacc tgccagatga agaggcaggg cgcccaagtg accaccgaga gaacagtgca
     1621 gctgaacgtg tcctacgctc cccagaccat caccatcttc cggaacggaa tcgccctgga
     1681 aatcctgcag aacaccagct acctgcctgt gctggaaggc caggctctga gactgctgtg
     1741 tgacgcccct agcaatcctc ctgctcacct gagctggttt cagggcagcc cagctctgaa
     1801 cgccacacct atcagcaaca ccggcatcct ggaactgaga agagtgcgga gcgctgagga
     1861 aggcggcttc acttgtagag cccagcatcc tctgggcttt ctgcagatct tcctgaacct
     1921 gagcgtgtac agcctgccac agctgctggg acctagctgt tcttgggaag ccgagggcct
     1981 gcactgccgg tgtagcttta gagctagacc cgctcctagc ctgtgttggc ggctggaaga
     2041 gaaacctctg gaaggcaata gcagccaggg cagcttcaaa gtgaacagca gctctgccgg
     2101 accttgggcc aatagctctc tgattctgca cggcggcctg agcagcgatc tgaaggtgtc
     2161 atgcaaggcc tggaacatct acggcagcca gtctggctct gtgctgctgc ttcagggaag
     2221 aagcaatctc ggcacaggcg tggtgcctgc tgctcttggc gcgccgggcg gcgacaagac
     2281 ccacacctgc cccccctgcc cagctccaga actgctggga gggccttccg tgttcctgtt
     2341 cccccccaag cccaaggaca ccctgatgat cagcaggacc cccgaggtga cctgcgtggt
     2401 ggtggccgtg tcccacgagg acccagaggt gaagttcaac tggtacgtgg acggcgtgga
     2461 ggtgcacaac gccaagacca agcccagaga ggagcagtac aacagcacct acagggtggt
     2521 gtccgtgctg accgtgctgc accaggactg gctgaacggc aaagaataca agtgcaaagt
     2581 ctccaacaag gccctggctg ccccaatcga aaagacaatc agcaaggcca agggccagcc
     2641 acgggagccc caggtgtaca ccctgccccc cagccgggag gagatgacca agaaccaggt
     2701 gtccctgacc tgtctggtga agggcttcta ccccagcgat atcgccgtgg agtgggagag
     2761 caacggccag cccgagaaca actacaagac caccccccca gtgctggaca gcgacggcag
     2821 cttcttcctg tacagcaagc tgaccgtgga caagtccagg tggcagcagg gcaacgtgtt
     2881 cagctgcagc gtgatgcacg aggccctgca caaccactac acccagaagt ccctgagcct
     2941 gagccccggc ggcagcggtc ttaatgatat tttcgaggct cagaagattg agtggcatga
     3001 acaccaccac catcaccact aaggccggcc tgtagtcact ggcacaacgc gttacccggt
     3061 aagccaatcg ggtatacacg gtcgtcatac tgcagacagg gttcttctac tttgcaagat
     3121 agtcttgagt agtaaaataa atagatagag aaaatcagcc tcgactgtgc cttctagttg
     3181 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3241 cactgtcctt tcctaataaa atgaggaaat tgcatcacaa cactcaaccc tatctcggtc
     3301 tattcttttg atttataagg gattttgccg atttyggcct attggttaaa aaatgagctg
     3361 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3481 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3661 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3721 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     3781 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3841 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3901 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3961 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4021 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4081 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4141 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4201 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4261 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4321 ggagacatcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4381 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     4441 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4501 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4561 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4681 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4741 gtcattacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4801 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4861 gccccagcgc tgcgatgata ccgcgagaac cacgctcacc ggctccggat ttatcagcaa
     4921 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4981 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5041 gcaacgttgt tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5101 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5161 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5221 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5281 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5341 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5401 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5461 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5521 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5581 cgacacggaa atgttgaata ctcatattct tcctttttca atattattga agcatttatc
     5641 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5701 gggtcagtgt tacaaccaat taaccaattc tgaacattat cgcgagccca tttatacctg
     5761 aatatggctc ataacacccc ttgctcatga ccaaaatccc ttaacgtgag ttacgcgcgc
     5821 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5881 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5941 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6001 gataccaaat actgttcttc tagtgtagcc gtagttagcc caccacttca agaactctgt
     6061 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6121 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6181 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6241 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6301 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6361 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6421 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6481 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6541 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6601 gaccgagcgc agcgagtcag tgagcgagga agcggaaggc gagagtaggg aactgccagg
     6661 catcaaacta agcagaaggc ccctgacgga tggccttttt gcgtttctac aaactctttc
     6721 tgtgttgtaa aacgacggcc agtcttaagc tcgggccccc tgggcggttc tgataacgag
     6781 taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt atggggggag
     6841 tttagggaaa gagcatttgt cagaatattt aagggcgcct gtcactttgc ttgatatatg
     6901 agaattattt aaccttataa atgagaaaaa agcaacgcac tttaaataag atacgttgct
     6961 ttttcgattg atgaacacct ataattaaac tattcatcta ttatttatga ttttttgtat
     7021 atacaatatt tctagtttgt taaagagaat taagaaaata aatctcgaaa ataataaagg
     7081 gaaaatcagt ttttgatatc aaaattatac atgtcaacga taatacaaaa tataatacaa
     7141 actataagat gttatcagta tttattatca tttagaataa attttgtgtc gcccttaatt
     7201 gtgagcggat aacaattacg agcttcatgc acagtggcgt tgactcaata ttggccatta
     7261 gccatattat tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg
     7321 ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg accgccatgt
     7381 t
//
