LOCUS       sequence-368838-        6484 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 6484)
  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 6484)
  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-ERMAP-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    351..462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     451..468
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(608..625)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(626..670)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1865..1884)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1897..1915)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1898..1915)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2125..2149)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2146..2349)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2175..2195)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2350..2729
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2896..2912
                     /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(3368..3385)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3368..3384)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3483..3500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3654..4242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3734..3753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4328..4347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4353..4372)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4414..4505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4506..4574,4575..5366)
                     /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(4724..4743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5419..5540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5430..5449)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5484..5503
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5670..6269)
                     /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(5753..5773)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6250..6269
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6288..6335)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6383..6484,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6397..6484,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6451..6470)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
      241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt gatgcaattt
      361 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      421 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gattttctct
      481 atctatttat tttactactc aagactatct tgcaaagtag aagaaccctg tctgcagtat
      541 gacgaccgtg tatacccgat tggcttaccg ggtaacgcgt tgtgccagtg actacaggcc
      601 ggccttagtg gtgatggtgg tggtgttcat gccactcaat cttctgagcc tcgaaaatat
      661 cattaagacc gctgccgccg gggctcaggc tcagggactt ctgggtgtag tggttgtgca
      721 gggcctcgtg catcacgctg cagctgaaca cgttgccctg ctgccacctg gacttgtcca
      781 cggtcagctt gctgtacagg aagaagctgc cgtcgctgtc cagcactggg ggggtggtct
      841 tgtagttgtt ctcgggctgg ccgttgctct cccactccac ggcgatatcg ctggggtaga
      901 agcccttcac cagacaggtc agggacacct ggttcttggt catctcctcc cggctggggg
      961 gcagggtgta cacctggggc tcccgtggct ggcccttggc cttgctgatt gtcttttcga
     1021 ttggggcagc cagggccttg ttggagactt tgcacttgta ttctttgccg ttcagccagt
     1081 cctggtgcag cacggtcagc acggacacca ccctgtaggt gctgttgtac tgctcctctc
     1141 tgggcttggt cttggcgttg tgcacctcca cgccgtccac gtaccagttg aacttcacct
     1201 ctgggtcctc gtgggacacg gccaccacca cgcaggtcac ctcgggggtc ctgctgatca
     1261 tcagggtgtc cttgggcttg ggggggaaca ggaacacgga aggccctccc agcagttctg
     1321 gagctgggca gggggggcag gtgtgggtct tgtcgccgcc cggcgcgcca gcagaaggag
     1381 acagagagcc cacagaagga gcggccactt gcaggatgac ggtgtcctct ttgctcaggt
     1441 tgcccacttg gatcagacat ctgtagctgc cctgatcttc cagccgcacg tccaggatct
     1501 gcagtgtcac agatccctct tgggcatccc gcacgagcac tgttctgccc ttgtactcag
     1561 gcatcaggtc ctcgtcctgg tccttgccat ctctgaagat gtgcacggcc tgagatctct
     1621 gaggaaatgg gcttctcagc cagcgaactt ctttaggcac ggtgccaggc cacagagaca
     1681 gaggacacaa cagttcggcg gttcctccga gcagggccac atgaaatttt ccagcgtcgc
     1741 cagcatgggc ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct
     1801 tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt
     1861 ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat
     1921 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt
     1981 gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc
     2041 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
     2101 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
     2161 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2221 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2281 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     2341 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     2401 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     2461 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     2521 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     2581 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     2641 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     2701 ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat
     2761 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
     2821 ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt
     2881 gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa
     2941 tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     3001 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     3061 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     3121 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     3181 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     3241 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
     3301 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
     3361 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
     3421 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
     3481 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3541 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3601 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3841 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
     4081 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4261 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
     4321 tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca
     4381 gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa
     4441 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4501 agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4561 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4621 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4681 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4741 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4801 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4861 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4921 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4981 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5041 cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5101 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5161 tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg
     5221 gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta
     5281 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5341 gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc
     5401 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5461 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5521 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     5581 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     5641 gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag
     5701 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     5761 aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc
     5821 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     5881 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     5941 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     6001 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     6061 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     6121 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     6181 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     6241 aggcgcaccg tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc gtattatact
     6301 atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg
     6361 atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     6421 acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta
     6481 gtca
//
