LOCUS       Exported                7966 bp DNA     circular SYN 28-SEP-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7966)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7966
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   1..190
                     /label=ITR
                     /note="color: #ffcc99"
     promoter        join(189..437,439..948,1032..1125)
                     /label=CAG promoter
                     /note="CAG promoter"
                     /note="This forward directional feature has 4 segments:
                        1:  189 ..  437 / #ffffff / CMV enhancer
                        2:  439 ..  716 / #ffffff / chicken beta-actin promoter
                        3:  717 ..  948 / #ffffff / chicken beta-actin precur...
                        4: 1032 .. 1125 / #ffffff / rabbit beta globin 
                     splice..."
     primer_bind     998..1016
                     /label=CMV_for
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     1140..1169
                     /label=Primer 1
                     /note="color: black; sequence: 
                     ccggaattcgtcaccatggtgagcaagggcgag; added: 2021-08-16"
     CDS             1152..1868
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1152 .. 1154 / #05fd14
                        2: 1155 .. 1157 / #05fd14 / 1a
                        3: 1158 .. 1868 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1875..1901
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     misc_feature    1875..1901
                     /label=HA-tag
                     /note="color: #a6acb3"
     primer_bind     complement(1877..1908)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     cgcggatccagcgtaatctgggacatcgtatggg; added: 2021-08-16"
     CDS             1908..1961
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 1958"
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     1963..1988
                     /label=SP 923 to 281
                     /note="color: black; sequence: 
                     tccccccgggaccatggcaagcagatggctg; added: 2021-09-10"
     CDS             1971..3725
                     /codon_start=1
                     /label=wt(Ms)NCLX
                     /note="color: #993366"
                     /translation="MASRWLALLWAPVFLCVALILETASGTGDPSTKAHGHIQFSAGSV
                     NQTAMADCRAVCGLNTSDRCDFVRRNPDCRSEAGYLDYLEGIFCYFPPNLLPLAITLYV
                     FWLLYLFLILGVTAAKFFCPNLSAISTNLKLSHNVAGVTFLAFGNGAPDIFSALVAFSD
                     PRTAGLAIGALFGAGVLVTTVVAGGITILHPFMAASRPFLRDIAFYMVAVFLTFTALYL
                     GRITLTWALGYLGLYVFYVVTVIICTWVYQRQRSRSLVHSISETPELLSESEEDQMSSN
                     TNSYDYGDEYRPLLLGRETTVQILIQALNPLDYRKWRTQSISWRVLKVVKLPVEFLLLL
                     TVPVVDPDKDDRNWKRPLNCLQLVISPLVLVLTLQSGVYGIYEIGGLLPVWAVVVIVGT
                     ALASVTFFATSNREPPRLHWLFAFLGFLTSALWINAAATEVVNILRSLGVIFRLSNTVL
                     GLTLLAWGNSIGDAFSDFTLARQGYPRMAFSACFGGIIFNILVGVGLGCLLQIIRNHVV
                     EVKLEPDGLLVWVLASALGLSLIFSLVSVPLQCFQLSKAYGLCLLLFYICFLVVVLLTE
                     FGVIHLKKA"
     primer_bind     complement(3707..3735)
                     /label=ASP 923 to 281
                     /note="color: black; sequence: 
                     actccgctcgagagccgccttcttcaggtgaatc; added: 2021-09-10"
     CDS             3732..3761
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     misc_feature    3783..4370
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(3868..3886)
                     /label=WPRE_rev
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(4253..4264)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4252"
                     /translation="IEGR"
     polyA_signal    4403..4627
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   4685..4866
                     /label=ITR
                     /note="color: #ffcc99"
     promoter        complement(4848..5177)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4862..4997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     rep_origin      complement(5423..6011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6182..7042)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6182 .. 6973 / #ccffcc
                        2: 6974 .. 7042 / #ccffcc / signal sequence
                     Cleavage site after base 6973"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7043..7147)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        7931..7949
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      241 tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca gtacatcaag
      301 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      361 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      421 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      481 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      541 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
      601 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      661 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      721 tcgctgcgac gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      781 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      841 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      901 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcggggct gtccgcgggg
      961 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     1021 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     1081 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc taagctagcg
     1141 aattcgtcac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1201 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1261 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1321 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1381 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1441 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1501 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1561 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1621 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1681 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1741 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1801 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1861 tgtacaaggc tagctaccca tacgatgtcc cagattacgc tggatccgag ggcagaggaa
     1921 gtcttctaac atgcggtgac gtggaggaga atcccggccc taccgggacc atggcaagca
     1981 gatggctggc tctgctctgg gctcctgtct tcctctgtgt ggctctgata ctggagacgg
     2041 cgtctgggac cggagaccca tccacaaaag cccatggaca catccagttt tcagctggaa
     2101 gtgtcaacca gactgccatg gcggattgcc gagccgtgtg tggcctgaac acatctgatc
     2161 gctgtgactt tgtcaggagg aatccggact gccgcagcga ggcgggctac ctggactacc
     2221 ttgagggcat cttctgctac ttccccccca acctcctccc tctggccatc accctctacg
     2281 ttttctggct gctttacctc tttctgatcc tgggagtcac cgcggccaag ttcttctgcc
     2341 ctaacctgtc agccatctcc actaacctca aactctccca caacgtggca ggtgtcacct
     2401 tcctggcctt tggaaatggc gctccagaca tcttcagtgc tttagtggct ttctcagacc
     2461 cacgtactgc cggcctggcc atcggggctc tgtttggtgc aggggtgctg gtcaccactg
     2521 tggtggctgg aggcatcacc atcctgcatc ccttcatggc tgcctccagg cccttcctca
     2581 gggacatcgc tttctacatg gtggctgtgt tcctaacctt cactgcactc tatctcggca
     2641 ggatcacgct gacgtgggcg ctgggttacc tgggcctcta cgtgttctac gtggtcacgg
     2701 tcatcatctg cacttgggtc taccaacggc agcgaagcag gtctctggtc cactccatat
     2761 cggagacacc agagttgctg tctgagtcag aggaggacca gatgtcttcc aacaccaaca
     2821 gctatgacta tggagatgaa taccggcctc tgttgctggg tcgggagacc actgtccaga
     2881 tcctgatcca agccctgaat cccttggact acaggaagtg gagaactcag tcgatatcct
     2941 ggagggtcct gaaggtagtc aagttgcctg tggagttctt gttgctgctc acagtaccgg
     3001 ttgtggaccc cgacaaggac gatcggaatt ggaaacggcc actcaactgt ctgcagctgg
     3061 tcatcagccc cctggtcctg gtcctgaccc tgcagtcggg ggtctatggc atctatgaga
     3121 tcggcggtct ccttcctgtc tgggctgtgg tggtgatcgt gggcacagcg ctggcttcag
     3181 tgaccttctt tgccacgtct aacagagaac cccctagact gcactggctc tttgctttcc
     3241 tgggtttcct gaccagtgcc ctgtggatca atgcagccgc cacagaggtg gtgaacatat
     3301 tacggtccct gggtgtgatc ttccgcctga gcaacaccgt cctagggctg accctcctgg
     3361 cctggggaaa cagcattgga gatgccttct cagatttcac gctggcccgc caaggatacc
     3421 ctcggatggc cttctccgcc tgtttcgggg gcatcatctt caacatcctg gttggtgtgg
     3481 ggctgggctg cttgctgcag atcatcagga accacgttgt ggaggtgaag ctggagccag
     3541 acggattact ggtgtgggtg ctggccagtg ccctgggcct cagcttgatc ttctccctgg
     3601 tctccgtgcc gcttcagtgt ttccagctca gcaaggctta cggcctctgc ctcctcctct
     3661 tctacatctg tttccttgtt gtggtcctgc tcacagagtt tggggtgatt cacctgaaga
     3721 aggcggctct cgagcagaaa ctcatctcag aagaggatct ggcagcataa aagcttatcg
     3781 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgtgc
     3841 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3901 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3961 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4021 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4081 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4141 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     4201 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4261 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4321 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     4381 tcgactcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4441 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4501 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4561 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4621 tctatggctt ctgaggcgga aagaaccagc tggggctcga ctagagcatg gctacgtaga
     4681 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     4741 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4801 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agagcttttt gcaaaagcct
     4861 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg
     4921 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     4981 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     5041 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     5101 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     5161 caccctaact gacacacatt ccacagctgc attaatgaat cggccaacgc gcggggagag
     5221 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5281 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5341 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5401 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5461 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5521 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5581 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5641 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5701 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5761 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5821 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5881 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5941 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6001 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6061 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6121 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6181 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6241 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6301 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6361 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6421 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6481 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6541 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6601 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6661 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6721 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6781 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6841 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6901 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6961 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7021 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7081 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7141 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7201 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     7261 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     7321 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     7381 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattcg acgctctccc
     7441 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     7501 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     7561 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     7621 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     7681 gccacgatgc gtccggcgta gaggatctgg cgatgaccct gctgattggt tcgctgacca
     7741 tttccgggtg cgggacggcg ttaccagaaa ctcagaaggt tcgtccaacc aaaccgactc
     7801 tgacggcagt ttacgagaga gatgataggg tctgcttcag taagccagat gctacacaat
     7861 taggcttgta catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca
     7921 tacacatacg atttaggtga cactatagaa tacacggaat taattc
//
