LOCUS       sequence-368522-        6571 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 6571)
  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 6571)
  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-CD7-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     236..256
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    374..485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     474..491
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(631..648)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(649..693)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1975..1994)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2007..2025)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2008..2025)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2235..2259)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2256..2459)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2285..2305)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2460..2839
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3006..3022
                     /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(3478..3495)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3478..3494)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3593..3610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3764..4352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3844..3863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4438..4457)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4463..4482)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4524..4615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4616..4684,4685..5476)
                     /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(4834..4853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5529..5650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5540..5559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5594..5613
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5780..6379)
                     /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(5863..5883)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6360..6379
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6398..6445)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6493..6571,1..251))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6507..6571,1..71)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(6561..6571,1..9))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
       61 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
      121 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
      181 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
      241 aactgacaca cattccacag aattaattcg cgttaaattt ttgttaaatc agctcatttt
      301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
      361 ggttgagtgt tgtgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
      421 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
      481 cacagtcgag gctgattttc tctatctatt tattttacta ctcaagacta tcttgcaaag
      541 tagaagaacc ctgtctgcag tatgacgacc gtgtataccc gattggctta ccgggtaacg
      601 cgttgtgcca gtgactacag gccggcctta gtggtgatgg tggtggtgtt catgccactc
      661 aatcttctga gcctcgaaaa tatcattaag accgctgccg ccggggctca ggctcaggga
      721 cttctgggtg tagtggttgt gcagggcctc gtgcatcacg ctgcagctga acacgttgcc
      781 ctgctgccac ctggacttgt ccacggtcag cttgctgtac aggaagaagc tgccgtcgct
      841 gtccagcact gggggggtgg tcttgtagtt gttctcgggc tggccgttgc tctcccactc
      901 cacggcgata tcgctggggt agaagccctt caccagacag gtcagggaca cctggttctt
      961 ggtcatctcc tcccggctgg ggggcagggt gtacacctgg ggctcccgtg gctggccctt
     1021 ggccttgctg attgtctttt cgattggggc agccagggcc ttgttggaga ctttgcactt
     1081 gtattctttg ccgttcagcc agtcctggtg cagcacggtc agcacggaca ccaccctgta
     1141 ggtgctgttg tactgctcct ctctgggctt ggtcttggcg ttgtgcacct ccacgccgtc
     1201 cacgtaccag ttgaacttca cctctgggtc ctcgtgggac acggccacca ccacgcaggt
     1261 cacctcgggg gtcctgctga tcatcagggt gtccttgggc ttggggggga acaggaacac
     1321 ggaaggccct cccagcagtt ctggagctgg gcaggggggg caggtgtggg tcttgtcgcc
     1381 gcccggcgcg ccaggcaaag ctgaagcagc aggaggatct ggaagggcac tagctgtctg
     1441 aggatcaggc agagcagagc ctgttggtgg agcaggaagt gcagaagctc taggaggggc
     1501 gtcgctacat ctgtgccagc cttgagactg ttcctctgtg accagcacca gggtgccaga
     1561 gccgtacacg ttcacttcgg tgatggcctg acaggtatat gtgccggtgt cgctcagctg
     1621 cagtctgtgc atggtgatgg tcaggttgtc ctggctgccg ctgaagtcga ttctgcctct
     1681 gaatcttctg tcggtggtgg gcaccacgcc atcttcgtag tagatgatgt cctgaggctg
     1741 gggtccaagt tgtctcagat agatgccccg caggccgcca gatgtagaac aggtgatatt
     1801 cacggaggcg cccacaggca ctgtggtaca gtgagggctc tgctgaactt cttgagcggc
     1861 ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct tcatggtggc
     1921 gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt ggacacctgt
     1981 ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat gtgtatgata
     2041 cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt gggcctatag
     2101 actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc aatgcaccgt
     2161 tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     2221 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac
     2281 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     2341 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     2401 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     2461 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     2521 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     2581 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     2641 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     2701 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     2761 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     2821 ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac
     2881 atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata
     2941 taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt gcatgaagct
     3001 cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa tgataataaa
     3061 tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat gtataatttt
     3121 gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta attctcttta
     3181 acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata gtttaattat
     3241 aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct tttttctcat
     3301 ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt aaatattctg
     3361 acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt
     3421 tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga gcttaagact
     3481 ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg
     3541 gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct tcctcgctca
     3601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac ggctacacta
     4201 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4381 ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg tcatgagcaa
     4441 ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca gaattggtta
     4501 attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4561 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag
     4621 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4681 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4741 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4801 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4861 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4921 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4981 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5041 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5101 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5161 agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5221 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5281 ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg gttctcgcgg
     5341 tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5401 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5461 gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     5521 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5581 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5641 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     5701 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     5761 cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     5821 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     5881 tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     5941 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     6001 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     6061 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     6121 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     6181 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     6241 tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga
     6301 acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg
     6361 tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc gtattatact atgccgatat
     6421 actatgccga tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg atatacaagc
     6481 tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6541 tagctcagag gcagaggcgg cctcggcctc t
//
