LOCUS       Exported                7278 bp ds-DNA     circular SYN 28-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    AAV-CMV-H2B-FusionRed.dna
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7278)
  AUTHORS   Christin Hong
  TITLE     Direct Submission
  JOURNAL   Exported Dec 7, 2020 from SnapGene Viewer 4.0.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7278)
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     LTR             6..138)
                     /label=ITR
                     /note="/vntifkey=19"
     misc_feature    139..184)
                     /label=additional AAV sequences
                     /label=additional\AAV\sequences
                     /note="/vntifkey=21"
     enhancer        315..694)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        695..898)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          1012..1487)
                     /label=-beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     primer_bind     complement(1482..1520))
                     /label=p1075
     primer_bind     1482..1520)
                     /label=p1057_c1008-ins-evrogenFP421_FOR
     primer_bind     complement(1488..1521))
                     /label=p1056_c1008-bb-3105_REV
     CDS             1543..1920)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1933..1942)
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1939..2637)
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     primer_bind     2626..2657)
                     /label=p1055_c1008-bb-3105.FOR
     primer_bind     complement(2634..2665))
                     /label=p1058_c1008-ins-evrogenFP421_REV
     primer_bind     complement(2655..2671))
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2765..3159)
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     misc_feature    3468..3513)
                     /label=additional AAV sequences
                     /label=additional\AAV\sequences
                     /note="/vntifkey=21"
     LTR             complement(3514..3646))
                     /label=ITR
                     /note="/vntifkey=19"
     rep_origin      4014..4527)
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4197..4226)
                     /label=p1019
     primer_bind     complement(4211..4246))
                     /label=p1020
     promoter        5326..5430)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5431..6291)
                     /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     5719..5754)
                     /label=p1009
     primer_bind     complement(5725..5761))
                     /label=p1010
     rep_origin      6462..7050)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taatgcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
       61 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      121 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
      181 catgctctag cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atgagcttgg
      241 cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
      301 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      361 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      421 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      481 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      541 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      601 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      661 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      721 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      781 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      841 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      901 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
      961 acaccgggac cgatccagcc tcccctcgaa gctgatcctg agaacttcag ggtgagtcta
     1021 tgggaccctt gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag
     1081 gggagaagta acagggtaca catattgacc aaatcagggt aattttgcat ttgtaatttt
     1141 aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct
     1201 aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa
     1261 agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt
     1321 ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca
     1381 gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct
     1441 aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg
     1501 tgagctcaag cttcgaattc tgcagtcgac ggtaccgcca ccatgccaga gccagcgaag
     1561 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
     1621 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
     1681 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt
     1741 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag
     1801 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag
     1861 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
     1921 gatccaccgg tcgccaccat ggtgagcgag ctgattaagg agaacatgcc catgaagctg
     1981 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
     2041 ccctacgagg gcacccagac catgagaatc aaggtcgtcg agggcggccc tctccccttc
     2101 gccttcgaca tcctggctac cagcttcatg tacggcagca gaaccttcat caagcaccct
     2161 ccgggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
     2221 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     2281 tgcctcatct acaacgtcaa ggttagaggg gtgaacttcc cagccaacgg ccctgtgatg
     2341 cagaagaaaa cactcggctg ggaggcctcc accgagacga tgtaccccgc tgacggcggc
     2401 ctggaaggcg catgtgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
     2461 cttgagacca catacagatc caagaaaccc gctacgaacc tcaagatgcc cggcgtctac
     2521 aacgtggacc acagactgga aagaatcaag gaggccgacg atgagaccta cgtcgagcag
     2581 cacgaggtgg ctgtggccag atactctact ggtggcgctg gtgatggagg taaatgagcg
     2641 gccgccaagc ttatcgatac cgtcgacctc gagggcccag atctaattca ccccaccagt
     2701 gcaggctgcc tatcagaaag tggtggctgg tgtggctaat gccctggccc acaagtatca
     2761 ctaagctcgc tttcttgctg tccaatttct attaaaggtt cctttgttcc ctaagtccaa
     2821 ctactaaact gggggatatt atgaagggcc ttgagcatct ggattctgcc taataaaaaa
     2881 catttatttt cattgcaatg atgtatttaa attatttctg aatattttac taaaaaggga
     2941 atgtgggagg tcagtgcatt taaaacataa agaaatgaag agctagttca aaccttggga
     3001 aaatacacta tatcttaaac tccatgaaag aaggtgaggc tgcaaacagc taatgcacat
     3061 tggcaacagc ccctgatgcc tatgccttat tcatccctca gaaaaggatt caagtagagg
     3121 cttgatttgg aggttaaagt tttgctatgc tgtattttac attacttatt gttttagctg
     3181 tcctcatgaa tgtcttttca ctacccattt gcttatcctg catctctcag ccttgactcc
     3241 actcagttct cttgcttaga gataccacct ttcccctgaa gtgttccttc catgttttac
     3301 ggcgagatgg tttctcctcg cctggccact cagccttagt tgtctctgtt gtcttataga
     3361 ggtctacttg aagaaggaaa aacagggggc atggtttgac tgtcctgtga gcccttcttc
     3421 cctgcctccc ccactcacag tgacccggaa tccctcgaca tctagagcat ggctacgtag
     3481 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3541 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3601 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag
     3661 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgattcc
     3721 gttgcaatgg ctggcggtaa tattgttctg gatattacca gcaaggccga tagtttgagt
     3781 tcttctactc aggcaagtga tgttattact aatcaaagaa gtattgcgac aacggttaat
     3841 ttgcgtgatg gacagactct tttactcggt ggcctcactg attataaaaa cacttctcag
     3901 gattctggcg taccgttcct gtctaaaatc cctttaatcg gcctcctgtt tagctcccgc
     3961 tctgattcta acgaggaaag cacgttatac gtgctcgtca aagcaaccat agtacgcgcc
     4021 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4081 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4141 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4201 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4261 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4321 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4381 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4441 ttttaacaaa atattaacgc ttacaattta aatatttgct tatacaatct tcctgttttt
     4501 ggggcttttc tgattatcaa ccggggtaca tatgattgac atgctagttt tacgattacc
     4561 gttcatcgat tctcttgttt gctccagact ctcaggcaat gacctgatag cctttgtaga
     4621 gacctctcaa aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat
     4681 catattgatg gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca
     4741 cattactcag gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt
     4801 gaaataaagg cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat
     4861 ttagctttat gctctgaggc tttattgctt aattttgcta attctttgcc ttgcctgtat
     4921 gatttattgg atgttggaat cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4981 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5041 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5101 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5161 catcaccgaa acgcgcgagc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5221 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5281 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6841 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6901 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6961 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7021 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7081 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7141 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7201 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7261 gcgcgttggc cgattcat
//
