LOCUS       sequence-368811-        6313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6313)
  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 6313)
  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-CCL4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2035..2054)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2067..2085)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2068..2085)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2295..2319)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2316..2519)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2345..2365)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2520..2899
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3066..3082
                     /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(3538..3555)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3538..3554)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3653..3670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3824..4412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3904..3923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4498..4517)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4523..4542)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4584..4675
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4676..4744,4745..5536)
                     /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(4894..4913)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5589..5710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5600..5619)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5654..5673
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5840..6313,1..126))
                     /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(5923..5943)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc attcagttcc aggtcgtaca cgtactcttg
     1741 cacccagctc tcgctaggat cggcgcacac ttgcttgctt ctcttggtct gaaacaccac
     1801 ggcgggctga gaacacaggc tgcttgtctc gtagtagtcc accacgaagt tccggggcag
     1861 ctttctggct gtgtagctga agcagcaggc ggtaggagga tcagatccca taggagcggc
     1921 ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct tcatggtggc
     1981 gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt ggacacctgt
     2041 ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat gtgtatgata
     2101 cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt gggcctatag
     2161 actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc aatgcaccgt
     2221 tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     2281 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac
     2341 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     2401 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     2461 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     2521 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     2581 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     2641 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     2701 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     2761 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     2821 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     2881 ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac
     2941 atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata
     3001 taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt gcatgaagct
     3061 cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa tgataataaa
     3121 tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat gtataatttt
     3181 gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta attctcttta
     3241 acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata gtttaattat
     3301 aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct tttttctcat
     3361 ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt aaatattctg
     3421 acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt
     3481 tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga gcttaagact
     3541 ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg
     3601 gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct tcctcgctca
     3661 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3721 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3781 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3841 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3901 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3961 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4021 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4081 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4141 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4201 cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac ggctacacta
     4261 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4321 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4381 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4441 ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg tcatgagcaa
     4501 ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca gaattggtta
     4561 attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4621 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag
     4681 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4741 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4801 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4861 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4921 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4981 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5041 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5101 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5161 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5221 agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5281 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5341 ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg gttctcgcgg
     5401 tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5461 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5521 gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     5581 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5641 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5701 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     5761 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     5821 cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     5881 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     5941 tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     6001 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     6061 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     6121 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     6181 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     6241 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     6301 tgacccgctc gat
//
