LOCUS       sequence-368860-        6508 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 6508)
  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 6508)
  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-IGSF23-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(140..187)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(235..564)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(303..322)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     549..569
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    687..798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     787..804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(944..961)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(962..1006)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2225..2244)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2257..2275)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2258..2275)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2485..2509)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2506..2709)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2535..2555)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2710..3089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3256..3272
                     /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(3728..3745)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3728..3744)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3843..3860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4014..4602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4094..4113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4688..4707)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4713..4732)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4774..4865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4866..4934,4935..5726)
                     /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(5084..5103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5779..5900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5790..5809)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5844..5863
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6030..6508,1..121))
                     /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(6113..6133)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
       61 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
      121 tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat
      181 tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
      241 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc
      301 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
      361 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      421 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      481 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      541 ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa
      601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      661 tagaccgaga tagggttgag tgttgtgatg caatttcctc attttattag gaaaggacag
      721 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      781 ggcaactaga aggcacagtc gaggctgatt ttctctatct atttatttta ctactcaaga
      841 ctatcttgca aagtagaaga accctgtctg cagtatgacg accgtgtata cccgattggc
      901 ttaccgggta acgcgttgtg ccagtgacta caggccggcc ttagtggtga tggtggtggt
      961 gttcatgcca ctcaatcttc tgagcctcga aaatatcatt aagaccgctg ccgccggggc
     1021 tcaggctcag ggacttctgg gtgtagtggt tgtgcagggc ctcgtgcatc acgctgcagc
     1081 tgaacacgtt gccctgctgc cacctggact tgtccacggt cagcttgctg tacaggaaga
     1141 agctgccgtc gctgtccagc actggggggg tggtcttgta gttgttctcg ggctggccgt
     1201 tgctctccca ctccacggcg atatcgctgg ggtagaagcc cttcaccaga caggtcaggg
     1261 acacctggtt cttggtcatc tcctcccggc tggggggcag ggtgtacacc tggggctccc
     1321 gtggctggcc cttggccttg ctgattgtct tttcgattgg ggcagccagg gccttgttgg
     1381 agactttgca cttgtattct ttgccgttca gccagtcctg gtgcagcacg gtcagcacgg
     1441 acaccaccct gtaggtgctg ttgtactgct cctctctggg cttggtcttg gcgttgtgca
     1501 cctccacgcc gtccacgtac cagttgaact tcacctctgg gtcctcgtgg gacacggcca
     1561 ccaccacgca ggtcacctcg ggggtcctgc tgatcatcag ggtgtccttg ggcttggggg
     1621 ggaacaggaa cacggaaggc cctcccagca gttctggagc tgggcagggg gggcaggtgt
     1681 gggtcttgtc gccgcccggc gcgccagagc cgccgctcag agacagtgta ggatcaggtt
     1741 ccataggctc ggcctcggta ggctgcatga taggcttagg caggctgatg gtcacaggct
     1801 cggacaccag ctgtttcttg ctgttggtgg cgatacacat gtaggtgccc agctgctcgc
     1861 aagacagcct tctgatgaac agcttctcgc ccatgccaca aggcactccg ctaaaggtcc
     1921 agctcagcac aggctcaggg tccattgtca cgacccactg caggatcacg ctgtagttca
     1981 gctcgctctg gatcacgcct ctgatggcgg caggaaaggt tttcaggggc atcagctgca
     2041 gaaccagatt agcaggaaaa tcgccgccag cggcgtcctt ttccagcata ggatcggttg
     2101 tggttgtggc ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct
     2161 tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt
     2221 ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat
     2281 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt
     2341 gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc
     2401 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
     2461 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     2521 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2581 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2641 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     2701 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     2761 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     2821 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     2881 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     2941 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3001 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3061 ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat
     3121 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
     3181 ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt
     3241 gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa
     3301 tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     3361 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     3421 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     3481 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     3541 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     3601 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     3661 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     3721 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     3781 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     3841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     4441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4621 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     4681 tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca
     4741 gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa
     4801 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4861 agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4921 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4981 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5041 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5101 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5161 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5221 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5281 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5341 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5401 cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5461 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5521 tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg
     5581 gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta
     5641 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5701 gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc
     5761 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5821 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5881 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     5941 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     6001 gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag
     6061 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     6121 aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc
     6181 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     6241 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     6301 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     6361 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     6421 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     6481 tgcagctcgg tgacccgctc gatgtggc
//
