LOCUS       sequence-368745-        6631 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 6631)
  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 6631)
  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-TNFRSF25-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..118
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(137..184)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(232..561)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      246..381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(300..319)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     546..566
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    684..795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     784..801
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(941..958)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(959..1003)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2345..2364)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2377..2395)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2378..2395)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2605..2629)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2626..2829)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2655..2675)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2830..3209
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3376..3392
                     /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(3848..3865)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3848..3864)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3963..3980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4134..4722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4214..4233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4808..4827)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4833..4852)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4894..4985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4986..5054,5055..5846)
                     /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(5204..5223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5899..6020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5910..5929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5964..5983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6150..6631,1..118))
                     /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(6233..6253)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
       61 cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg
      121 tttagttcct caccttgtcg tattatacta tgccgatata ctatgccgat gattaattgt
      181 caacacgtgc tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa
      241 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc
      301 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      361 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      421 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      481 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      541 tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt ttgttaaatc
      601 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      661 accgagatag ggttgagtgt tgtgatgcaa tttcctcatt ttattaggaa aggacagtgg
      721 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
      781 aactagaagg cacagtcgag gctgattttc tctatctatt tattttacta ctcaagacta
      841 tcttgcaaag tagaagaacc ctgtctgcag tatgacgacc gtgtataccc gattggctta
      901 ccgggtaacg cgttgtgcca gtgactacag gccggcctta gtggtgatgg tggtggtgtt
      961 catgccactc aatcttctga gcctcgaaaa tatcattaag accgctgccg ccggggctca
     1021 ggctcaggga cttctgggtg tagtggttgt gcagggcctc gtgcatcacg ctgcagctga
     1081 acacgttgcc ctgctgccac ctggacttgt ccacggtcag cttgctgtac aggaagaagc
     1141 tgccgtcgct gtccagcact gggggggtgg tcttgtagtt gttctcgggc tggccgttgc
     1201 tctcccactc cacggcgata tcgctggggt agaagccctt caccagacag gtcagggaca
     1261 cctggttctt ggtcatctcc tcccggctgg ggggcagggt gtacacctgg ggctcccgtg
     1321 gctggccctt ggccttgctg attgtctttt cgattggggc agccagggcc ttgttggaga
     1381 ctttgcactt gtattctttg ccgttcagcc agtcctggtg cagcacggtc agcacggaca
     1441 ccaccctgta ggtgctgttg tactgctcct ctctgggctt ggtcttggcg ttgtgcacct
     1501 ccacgccgtc cacgtaccag ttgaacttca cctctgggtc ctcgtgggac acggccacca
     1561 ccacgcaggt cacctcgggg gtcctgctga tcatcagggt gtccttgggc ttggggggga
     1621 acaggaacac ggaaggccct cccagcagtt ctggagctgg gcaggggggg caggtgtggg
     1681 tcttgtcgcc gcccggcgcg ccttgtcgcc agccacaaac agcggcacat ctctcaggac
     1741 agctgcccag tgtagatgta ggacaggaca cgcagccgtc gccgtgttca taaaagccag
     1801 gcagacaggt gccgcagtcg gtgtctcttc tgctacacag cagtctggtg tgtctgtgca
     1861 gggcgccaca atccagacaa ggctggcagt agaaggggct gctagacaca cactgggaca
     1921 cctggcattc cacgaaccat ccaggcttgc atccgcatct ggtatcggcc acggcagagc
     1981 aattttccag ggccacctga gaagcctgtt catcacaggc ctggcatctg gcacactcgc
     2041 tattgtggtg gttctcccag gccaggaagg tatcttgagg gcacacgagg catgtgctat
     2101 tgccgcaggg ctctgtacaa ggggccttca ggtagtgtcc ggcaggacat cctctgcagc
     2161 agaacaggcc gatcttcttg tggaagtcgc cggcacagtc acatctagga gatctggttc
     2221 caccttgggc ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct
     2281 tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt
     2341 ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat
     2401 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt
     2461 gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc
     2521 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
     2581 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     2641 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2701 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2761 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     2821 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     2881 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     2941 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3001 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3061 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3121 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3181 ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat
     3241 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
     3301 ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt
     3361 gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa
     3421 tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     3481 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     3541 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     3601 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     3661 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     3721 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     3781 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     3841 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     3901 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     3961 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4021 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4081 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4141 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4201 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4261 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4321 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4381 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4441 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4501 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     4561 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4621 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4681 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4741 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     4801 tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca
     4861 gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa
     4921 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4981 agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5041 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5101 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5161 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5221 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5281 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5341 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5401 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5461 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5521 cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5581 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5641 tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg
     5701 gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta
     5761 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5821 gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc
     5881 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5941 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     6001 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     6061 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     6121 gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag
     6181 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     6241 aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc
     6301 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     6361 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     6421 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     6481 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     6541 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     6601 tgcagctcgg tgacccgctc gatgtggcgg t
//
