LOCUS       Exported                5413 bp ds-DNA     circular SYN 17-APR-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5413)
  AUTHORS   Leonard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 17, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5413
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        228..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        608..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             900..977
                     /codon_start=1
                     /label=CD4 Signal Sequence
                     /translation="MCRAISLRRLLLLLLQLSQLLAVTQG"
     CDS             978..1043
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1044..1367
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             1368..1547
                     /codon_start=1
                     /label=Linker
                     /translation="SAGGSAGGSAGGSAGGSAGGPRAQASNSAVDGTAGPGSTGSRGGS
                     GGSGGSGGSGGSGGS"
     CDS             1554..1616
                     /codon_start=1
                     /label=CD28 TMD
                     /translation="LVVVAGVLFCYGLLVTVALCV"
     CDS             1617..1634
                     /codon_start=1
                     /label=Linker
                     /translation="GGGSGG"
     CDS             1641..1991
                     /codon_start=1
                     /label=NTEVp-118
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQSLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQT"
     misc_feature    1860..1862
                     /label=Mutation
     polyA_signal    2045..2269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2315..2743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2813..2948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3082..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3252..3268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3276..3292
                     /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)."
     promoter        complement(3300..3330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3345..3366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3654..4239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4410..5270)
                     /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"
     promoter        complement(5271..5375)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
       61 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      121 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      181 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      841 ctggcttatc gaaattaata cgactcacta tagggagtcc caagctggct agcgccatca
      901 tgtgccgagc catctctctt aggcgcttgc tgctgctgct gctgcagctg tcacaactcc
      961 tagctgtcac tcaaggggat tacaaggatc acgatgggga ttacaaagat catgacattg
     1021 actacaagga tgacgatgat aagatgggag tgcaggtgga aaccatctcc ccaggagacg
     1081 ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag
     1141 atggaaagaa atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
     1201 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
     1261 ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc
     1321 caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaagt gctggtggta
     1381 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtcct cgagctcaag
     1441 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agaggcggat
     1501 ctggaggctc tgggggttca ggcggatcag gaggcagtgg cgggtcaacc ggtcttgtcg
     1561 tcgttgcagg ggtgctcttc tgttatggtc tgttggtgac agtcgcactg tgtgtaggag
     1621 gcggtagcgg aggcggcgcc gagagcttgt ttaaggggcc gcgtgattac aacccgatat
     1681 cgagcaccat ttgtcatttg acgaatgaat ctgatgggca cacaacatcg ttgtatggta
     1741 ttggatttgg tcccttcatc attacaaaca agcacttgtt tagaagaaat aatggaacac
     1801 tgttggtcca atcactacat ggtgtattca aggtcaagaa caccacgact ttgcaacaaa
     1861 gcctcattga tgggagggac atgataatta ttcgcatgcc taaggatttc ccaccatttc
     1921 ctcaaaagct gaaatttaga gagccacaaa gggaagagcg catatgtctt gtgacaacca
     1981 acttccaaac ttaggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     2041 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2101 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2161 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2221 gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2281 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2341 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2401 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2461 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2521 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2581 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2641 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2701 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2761 gtcagttagg ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2821 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2881 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2941 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggaggttg
     3001 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3061 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3121 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3181 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3241 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3301 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3361 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3421 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     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 gcctaactac
     4081 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     4201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4261 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4321 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4381 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4441 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4501 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc
     4561 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4621 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4681 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4741 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4801 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4861 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4921 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4981 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5041 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5101 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5161 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5221 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5281 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5341 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5401 gacgtcgacg gat
//
