LOCUS       Exported                6819 bp ds-DNA     circular SYN 22-MAY-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2019 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6819
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        236..852
                     /label=CMV_immearly_promoter
                     /note="color: #ffffff"
     primer_bind     769..789
                     /label=CMV_fwd_primer
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     promoter        863..881
                     /label=T7_promoter
                     /note="color: #ffffff"
     misc_feature    935..940
                     /label=KOZAK
                     /note="color: #ffffff"
     CDS             941..1039
                     /codon_start=1
                     /label=TOM20 signal peptide
                     /note="color: #993366"
                     /translation="MVGRNSAIAAGVCGALFIGYCIYFDRKRRSDPN"
     primer_bind     1040..1083
                     /label=Fw AgeI jRCaMP1b for OMM
                     /note="color: black; sequence: 
                     GCTGATGCGACCGGTcatcatcatcatcatcatggtatggctagcatgactgg"
     CDS             1040..1045
                     /codon_start=1
                     /label=linker
                     /note="color: #c0c0c0"
                     /translation="TG"
     CDS             1046..1063
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             1067..1099
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /note="color: #cc99b2"
                     /translation="MASMTGGQQMG"
     CDS             1103..1126
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /note="This feature has 2 segments:
                        1: 1103 .. 1111 / #cc99b2
                        2: 1112 .. 1126 / #cc99b2 / enterokinase site
                     Cleavage site after base 1126"
                     /translation="DLYDDDDK"
     CDS             1127..2362
                     /codon_start=1
                     /label=jRCaMP1b
                     /note="color: #ff0000"
                     /translation="DLATMVDSSRRKWNKWGHAVRAIGRLSSANNTEMMYPADGGLRGY
                     THMALKVDGGGHLSCSFVTTYRSKKTVGNIKMPAIHYVSHRLERLEESDNEMFVVQREH
                     AVAKFVGLGGGGGTGGSMNSLIKENMRMKVVLEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     ITVMQDTSLEDGCLVYHVQVRGVNFPSNGAVMQKKTKGWEPTDSQLTEEQIAEFKEAFS
                     LFDKDGDGTITTKEMGTVMRSLGQNPTEAELQDMINEVDADGDGTIDFPEFLIMMAGKM
                     KYTDSEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADSDGDGQ
                     VNYEEFVQMMTAK"
     primer_bind     complement(2410..2427)
                     /label=BGH rev primer
                     /note="color: black; sequence: TAGAAGGCACAGTCGAGG"
     polyA_site      2416..2640
                     /label=BGH pA
                     /note="color: #a6acb3"
     rep_origin      2703..3009
                     /label=f1_origin
                     /note="color: #ffff00"
     primer_bind     complement(3123..3143)
                     /label=pBABE_3_primer
                     /note="color: black; sequence: ACCCTAACTGACACACATTCC"
     promoter        3261..3463
                     /label=SV40_promoter
                     /note="color: #ffffff"
     primer_bind     3370..3389
                     /label=SV40pro_F_primer
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     CDS             3515..4315
                     /codon_start=1
                     /label=Neo(R)
                     /note="color: #993366"
                     /translation="SFRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      4497..4613
                     /label=SV40_PA_terminator
                     /note="color: #ffffff"
     primer_bind     4583..4602
                     /label=EBV_rev_primer
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     primer_bind     complement(4658..4676)
                     /label=M13_reverse_primer
                     /note="color: black; sequence: GGAAACAGCTATGACCATG"
     primer_bind     complement(4675..4697)
                     /label=M13_pUC_rev_primer
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     rep_origin      complement(5049..5668)
                     /direction=LEFT
                     /label=pBR322_origin
                     /note="color: #ffff00"
     CDS             complement(5823..6683)
                     /codon_start=1
                     /label=Amp(R)
                     /note="color: #ccffcc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6725..6753)
                     /label=AmpR_promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggtgggaa gaaactctgc
      961 tatcgccgct ggcgtttgtg gcgctctgtt catcggctac tgcatctact tcgacagaaa
     1021 gagaagaagc gaccccaaca ccggtcatca tcatcatcat catggtatgg ctagcatgac
     1081 tggtggacag caaatgggtc gggatctgta cgacgatgac gataaggatc tcgcaacaat
     1141 ggtcgactca tcgcgacgta agtggaataa gtggggtcac gcagtcagag ctataggtcg
     1201 gctgagctca gcgaacaaca ccgaaatgat gtacccagcg gatggtggtc tgcgtggtta
     1261 cactcacatg gcgctgaaag ttgatggcgg cggtcacctg tcctgttctt tcgtgaccac
     1321 ctaccgctcc aaaaagactg tcggcaacat taagatgcct gccattcatt acgtcagcca
     1381 ccgtctggag cgcctggagg agagcgataa cgaaatgttt gtcgtacagc gtgaacacgc
     1441 agttgccaag tttgtgggcc tgggtggtgg cggcggtacc ggagggagca tgaactccct
     1501 gatcaaggag aacatgcgta tgaaagtggt tctggaaggc tccgtaaacg gccaccagtt
     1561 caaatgcact ggtgaaggcg aaggcaaccc gtatatgggc acccagacta tgcgtatcaa
     1621 agtgatcgag ggtggtccgc tgccgtttgc gttcgacatc ctggcgacgt cctttatgta
     1681 tggctcccgt accttcatca aatatccgaa aggcatcccg gatttcttta agcagtcctt
     1741 cccggaaggt tttacctggg aacgtgtgac ccgttacgaa gacggcggcg taattaccgt
     1801 tatgcaagac acgtctctgg aggatggctg cctggtgtat cacgtgcagg ttcgcggtgt
     1861 gaacttcccg agcaatggtg ctgtaatgca aaagaaaacc aaaggttggg agcctacgga
     1921 ctcccaactg actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa
     1981 ggacggggat gggacaataa caaccaagga gatggggacg gtgatgcggt ctctggggca
     2041 gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg
     2101 cacaatcgac ttccctgagt tcctgattat gatggcaggc aaaatgaaat acacagacag
     2161 tgaagaagaa attagagaag cgttcggcgt gtttgataag gatggcaatg gctacatcag
     2221 tgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag atgaagaggt
     2281 tgatgaaatg atcagggaag cagacagcga tggggatggt caggtaaact acgaagagtt
     2341 tgtacaaatg atgacagcga agtaggcggc cgctcgagtc tagagggccc gtttaaaccc
     2401 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2461 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2521 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2581 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2641 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     2701 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2761 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2821 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2881 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2941 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3001 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3061 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3121 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3181 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3301 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3361 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3421 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3481 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3541 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3601 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3661 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     3721 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3781 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3841 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3901 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3961 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4021 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4081 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4141 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4201 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4261 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4321 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4381 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4441 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4501 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4561 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4621 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4681 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6781 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
