LOCUS       Exported                4855 bp DNA     circular SYN 28-FEB-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4855)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 28, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
COMMENT     EE128 NLS-LgBit-FLAG-CaM from 1 to 4855
            EE40 LgBiT-15aa linker-FLAG-CaM (1) from 1 to 4801
FEATURES             Location/Qualifiers
     source          1..4855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..130
                     /locus_tag="ITR(1)"
                     /label=ITR
                     /label=ITR(1)
                     /note="color: #00ffff"
     misc_feature    151..708
                     /locus_tag="CMV promoter"
                     /label=CMV promoter
                     /note="color: #00ffff"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    753..769
                     /locus_tag="T7 promoter primer"
                     /label=T7 promoter primer
                     /note="color: #00ff00"
     misc_feature    785..786
                     /locus_tag="NotI"
                     /label=NotI
                     /note="color: #ff8040"
     misc_feature    791..793
                     /locus_tag="Kozak consensus "
                     /label=Kozak consensus
                     
                     /note="color: #ffd92b"
     misc_feature    791..793
                     /locus_tag="start codon"
                     /label=start codon
                     /note="color: #00ffff"
     misc_feature    794..847
                     /locus_tag="strong NLS"
                     /label=strong NLS
                     /note="color: #ffff66"
     CDS             827..847
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     misc_feature    848..1324
                     /locus_tag="Nanobit 11S (CO)"
                     /label=Nanobit 11S (CO)
                     /note="color: #9eafd2"
     misc_feature    1325..1378
                     /locus_tag="flexible linker"
                     /label=flexible linker
                     /note="color: #85dae9"
     misc_feature    1379..1402
                     /locus_tag="Flag"
                     /label=Flag
                     /note="color: #b4abac"
     CDS             1379..1402
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /note="color: #cc99b2"
                     /translation="DYKDDDDK"
     misc_feature    1403..1843
                     /locus_tag="Calmodulin"
                     /label=Calmodulin
                     /note="color: #ff8000"
     misc_feature    1844..1846
                     /locus_tag="END"
                     /label=END
                     /note="color: #00ffff"
     misc_feature    1847..1852
                     /locus_tag="EcoRI"
                     /label=EcoRI
                     /note="color: #46ff6e"
     misc_feature    1859..2090
                     /locus_tag="PolyA"
                     /label=PolyA
                     /note="color: #ff2549"
     polyA_signal    1883..2090
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    2119..2259
                     /locus_tag="ITR"
                     /label=ITR
                     /note="color: #00ffff"
     repeat_region   2119..2259
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      2333..2788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3070..3174
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3175..4035
                     /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: 3175 .. 3243 / #ccffcc / signal sequence
                        2: 3244 .. 4035 / #ccffcc
                     Cleavage site after base 3243"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4206..4794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atgaagcgga ctgctgatgg cagtgaattt gagagcccca aaaagaagag
      841 gaaagtggtc ttcactctgg aagatttcgt aggtgactgg gagcaaacag cagcatacaa
      901 tcttgatcaa gtgttggagc aagggggggt aagctctctc ctccaaaact tggctgtctc
      961 agttactcca attcaacgaa ttgtacggtc aggggaaaat gcactcaaga ttgacattca
     1021 tgtcatcatt ccttatgagg gtctgagtgc cgatcagatg gcccagatcg aggaggtttt
     1081 taaggtagtg tatcccgtgg atgatcatca ctttaaggta attttgccct acggaactct
     1141 ggtaattgac ggagtgaccc ccaatatgct caattatttt ggtaggccct atgaaggtat
     1201 tgctgtcttc gacggaaaaa aaataacagt tactgggaca ctctggaatg gtaataagat
     1261 tatagacgaa cgcttgatca ctcctgacgg cagtatgttg ttccgcgtta ccataaatgg
     1321 gtctgggaac ggaaatggca atggtaacgg gaatggaaac ggcaatggta acgggaatga
     1381 ctacaaggat gacgacgata aggaccaact gactgaagag cagatcgcag aatttaaaga
     1441 ggctttctcc ctatttgaca aggacgggga tgggacaata acaaccaagg agctggggac
     1501 ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca tgatcaatga
     1561 agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa tgatggcaag
     1621 aaaaatgaaa tacagggaca cggaagaaga aattagagaa gcgttcggtg tgtttgataa
     1681 ggatggcaat ggctacatca gtgcagcaga gcttcgccac gtgatgacaa accttggaga
     1741 gaagttaaca gatgaagagg ttgatgaaat gatcagggaa gcagacatcg atggggatgg
     1801 tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aagtaggaat tcctcgagct
     1861 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1921 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1981 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2041 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga ggtaaccacg
     2101 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2161 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2221 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2281 tccttacgca tctgtgcggt attcacaccg catacgtcaa agcaaccata gtacgcgccc
     2341 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2401 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2461 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2521 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2581 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2641 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2701 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2761 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     2821 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2881 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2941 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3001 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3061 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3121 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3181 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3241 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3301 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3361 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3421 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3481 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3541 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3601 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3661 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3721 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3781 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3841 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt
     3901 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3961 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4021 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4081 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4141 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4201 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4261 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4321 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4381 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4441 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4501 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4561 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4621 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4681 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4741 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4801 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
