LOCUS       sequence-368752-        6850 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of mCherry 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 6850)
  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 6850)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-mCherry-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6850
                     /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..975)
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             complement(976..1020)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(1711..1734)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      2451..2460
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2572..2591)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2604..2622)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2605..2622)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2832..2856)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2853..3056)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2882..2902)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3057..3436
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3603..3619
                     /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(4075..4092)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4075..4091)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4190..4207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4361..4949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4441..4460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5035..5054)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5060..5079)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        5121..5212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5213..5281,5282..6073)
                     /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(5431..5450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6126..6247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6137..6156)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6191..6210
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6377..6850,1..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(6460..6480)
                     /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 cggccttaat ggtgatggtg
      961 atggtgatgg tgatgttcgt gccattcgat tttctgagcc tcgaagatgt cgttcagtcc
     1021 tccgccgccg gggctcaggc tcagggactt ctgggtgtag tggttgtgca gggcctcgtg
     1081 catcacgctg cagctgaaca cgttgccctg ctgccacctg gacttgtcca cggtcagctt
     1141 gctgtacagg aagaagctgc cgtcgctgtc cagcactggg ggggtggtct tgtagttgtt
     1201 ctcgggctgg ccgttgctct cccactccac ggcgatatcg ctggggtaga agcccttcac
     1261 cagacaggtc agggacacct ggttcttggt catctcctcc cggctggggg gcagggtgta
     1321 cacctggggc tcccgtggct ggcccttggc cttgctgatt gtcttttcga ttggggcagc
     1381 cagggccttg ttggagactt tgcacttgta ttctttgccg ttcagccagt cctggtgcag
     1441 cacggtcagc acggacacca ccctgtaggt gctgttgtac tgctcctctc tgggcttggt
     1501 cttggcgttg tgcacctcca cgccgtccac gtaccagttg aacttcacct ctgggtcctc
     1561 gtgggacacg gccaccacca cgcaggtcac ctcgggggtc ctgctgatca tcagggtgtc
     1621 cttgggcttg ggggggaaca ggaacacgga aggccctccc agcagttctg gagctgggca
     1681 gggggggcag gtgtgggtct tgtcgctccc aggcccttga aacagaactt ccaaccccgg
     1741 cgcgcctttg tacagttcgt ccataccacc ggtagagtga cgaccttcag cacgttcgta
     1801 ctgttcaacg atggtgtagt cttcgttgtg agaggtgatg tccagtttga tgttaacgtt
     1861 gtaagcaccc ggcagctgaa ccggtttttt agctttgtag gtggttttaa cttcagcgtc
     1921 gtagtgacca ccgtctttca gtttcagacg ctgtttgatt tcacctttca gagcaccgtc
     1981 ttccgggtac atacgttcag aagaagcttc ccaacccatg gtttttttct gcataaccgg
     2041 accgtcagac gggaagttgg taccacgcag tttaactttg tagatgaatt caccgtcctg
     2101 cagagaagag tcctgggtaa cggtaacaac accaccgtct tcgaagttca taacacgttc
     2161 ccatttgaaa ccttccggga aagacagttt caggtagtcc gggatgtcag ccgggtgttt
     2221 aacgtaagct ttagaaccgt acatgaactg cggagacagg atgtcccaag cgaacggcag
     2281 cggaccacct ttggtaactt tcagtttagc ggtctgggta ccttcgtacg gacgaccttc
     2341 accttcacct tcgatttcga attcgtgacc gttaacagaa ccttccatgt gaactttgaa
     2401 acgcatgaat tctttgatga tagccatgtt gtcttcttca cctttagaaa ccatggcggc
     2461 cgcagcgaac accaggcaga agatgtagct cagggcaatg atggtcttca tggtggcgct
     2521 agcaggcgcg ttttcgatag aaccgaggtg cagttggacc tgggagtgga cacctgtgga
     2581 gagaaaggca aagtggatgt tattctatag tgtcacctaa atcgtatgtg tatgatacat
     2641 aaggttatgt attaattgta gccgcgttct aacgaagcca agggggtggg cctatagact
     2701 ctataggcgg tacttacgtc actcttggca cggggaatcc gcgttccaat gcaccgttcc
     2761 cggccgcgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg
     2821 cgtctccagg cgatctgacg gttcactaaa cgagctctgc ttatatagac ctcccaccgt
     2881 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
     2941 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     3001 ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg
     3061 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
     3121 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
     3181 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     3241 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     3301 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     3361 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     3421 gtcaataatc aatgccaaca tggcggtcat attggacatg agccaatata aatgtacata
     3481 ttatgatata gatacaacgt atgcaatggc caatagccaa tattgattta tgctatataa
     3541 ccaatgaata atatggctaa tggccaatat tgagtcaacg ccactgtgca tgaagctcgt
     3601 aattgttatc cgctcacaat taagggcgac acaaaattta ttctaaatga taataaatac
     3661 tgataacatc ttatagtttg tattatattt tgtattatcg ttgacatgta taattttgat
     3721 atcaaaaact gattttccct ttattatttt cgagatttat tttcttaatt ctctttaaca
     3781 aactagaaat attgtatata caaaaaatca taaataatag atgaatagtt taattatagg
     3841 tgttcatcaa tcgaaaaagc aacgtatctt atttaaagtg cgttgctttt ttctcattta
     3901 taaggttaaa taattctcat atatcaagca aagtgacagg cgcccttaaa tattctgaca
     3961 aatgctcttt ccctaaactc cccccataaa aaaacccgcc gaagcgggtt tttacgttat
     4021 ttgcggatta acgattactc gttatcagaa ccgcccaggg ggcccgagct taagactggc
     4081 cgtcgtttta caacacagaa agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc
     4141 ttctgcttag tttgatgcct ggcagttccc tactctcgcc ttccgcttcc tcgctcactg
     4201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4741 ggtatgtagg cggtgctaca gagttcttga agtggtgggc taactacggc tacactagaa
     4801 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4981 acgctcagtg gaacgacgcg cgcgtaactc acgttaaggg attttggtca tgagcaaggg
     5041 gtgttatgag ccatattcag gtataaatgg gctcgcgata atgttcagaa ttggttaatt
     5101 ggttgtaaca ctgaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5161 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga atatgagtat
     5221 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5281 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5341 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5401 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5461 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5521 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5581 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5641 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5701 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5761 gatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5821 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5881 tccggctggc tggtttattg ctgataaatc cggagccggt gagcgtggtt ctcgcggtat
     5941 catcgcagcg ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6001 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6061 taagcattgg taatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     6121 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6181 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6241 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     6301 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     6361 aatctcgtag cacgtgtcag gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt
     6421 cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgc
     6481 ggggcgcgga tgtctccagg aaggcgggca ccccggcgcg ctcggccgcc tccactccgg
     6541 ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg
     6601 ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc atctgttgct
     6661 gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg
     6721 cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga
     6781 cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga
     6841 cccgctcgat
//
