LOCUS       Exported                7197 bp ds-DNA     circular SYN 22-MAY-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Mito4xGCaMP6f
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2019 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7197
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    907..912
                     /label=KOZAK
                     /note="color: #ffffff"
     CDS             913..1008
                     /codon_start=1
                     /label=#1 COX8 sig pept
                     /note="color: #993366"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP"
     CDS             1009..1104
                     /codon_start=1
                     /label=#2 COX8 sig pept
                     /note="color: #993366"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP"
     CDS             1105..1200
                     /codon_start=1
                     /label=#3 COX8 sig pept
                     /note="color: #993366"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP"
     CDS             1201..1296
                     /codon_start=1
                     /label=#4 COX8 sig pept
                     /note="color: #993366"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAKIHSLGDP"
     CDS             1297..1323
                     /codon_start=1
                     /label=linker
                     /note="color: #c0c0c0"
                     /translation="RSGSAKDPT"
     CDS             1324..2673
                     /codon_start=1
                     /label=GCaMP6f
                     /note="color: #339966"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             1333..1350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             1390..1413
                     /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: 1390 .. 1398 / #cc99b2
                        2: 1399 .. 1413 / #cc99b2 / enterokinase site
                     Cleavage site after base 1413"
                     /translation="DLYDDDDK"
     promoter        complement(2749..2767)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    2793..3017
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3063..3491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3505..3835
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3686..3821
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3902..4696
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4870..4991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5040..5056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5064..5080
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5088..5118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5088 .. 5094 / #ffffff / -10
                        2: 5095 .. 5112 / #ffffff
                        3: 5113 .. 5118 / #ffffff / -35"
     protein_bind    5133..5154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5442..6030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6201..7061)
                     /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: 6201 .. 6992 / #ccffcc
                        2: 6993 .. 7061 / #ccffcc / signal sequence
                     Cleavage site after base 6992"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7062..7166)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ggatccaccg ccatgagcgt gctgacacct ctgctgctga gaggcctgac aggctctgct
      961 agaaggctgc ctgtgcctag agccaagatc cactctctgg gcgaccctat gtctgtgctg
     1021 actccactgc tcctgcgggg actgactgga tctgctcgta gactgccagt gccacgggct
     1081 aagattcaca gcctgggcga tccaatgagt gtgcttaccc cactcctgct cagaggactg
     1141 accggaagtg ctagacgcct gccagttcca agggccaaaa tccatagtct gggagatccc
     1201 atgtccgtgt tgacacccct cttgctgagg ggacttaccg gctcagctcg gagactccct
     1261 gttccaagag caaagattca ttccctcggc gacccaagat ctggcagcgc taaagaccct
     1321 accatgggta gccaccacca ccatcaccac ggcatggctt ctatgacagg cggccagcag
     1381 atgggccgcg acctgtacga tgacgacgat aaggacctgg ccacaatggt ggacagcagc
     1441 aggcggaagt ggaacaagac aggccacgcc gttagagcca tcggcagact gtctagcctg
     1501 gaaaacgtgt acatcaaggc cgacaagcag aagaacggca tcaaggctaa cttcaagatc
     1561 cggcacaaca tcgaggacgg cggagtgcag ctggcttacc actaccagca gaacacccct
     1621 atcggcgacg gaccagttct gctgcctgac aaccactacc tgagcgtgca gagcaagctg
     1681 agcaaggacc ccaacgagaa gagggaccac atggtgctgc tggaatttgt gaccgccgct
     1741 ggcatcacac tcggcatgga cgaactgtac aaaggcggca caggcggctc tatggtgtcc
     1801 aagggcgaag aactgttcac cggcgtggtg cccattctgg tggaactgga tggggatgtg
     1861 aacggccaca agttcagcgt cagcggagaa ggcgaaggcg acgccacata cggaaagctg
     1921 accctgaagt tcatctgcac caccggcaaa ctgcccgtgc cttggcctac actggtcacc
     1981 acactgacat acggcgtgca gtgcttcagc agataccccg accatatgaa gcagcacgac
     2041 ttcttcaaga gcgccatgcc tgagggctac atccaagaga gaaccatctt ctttaaggac
     2101 gacggcaact acaagaccag ggccgaagtg aagttcgagg gcgacaccct ggtcaacaga
     2161 atcgagctga agggcatcga cttcaaagag gacggcaaca tcctgggcca caagctcgag
     2221 tacaacctgc ctgaccagct gaccgaggaa cagatcgccg agttcaaaga agagttcagc
     2281 ctgttcgaca aggacggcga cggcacaatc accaccaaag aactgggcac cgtgatgaga
     2341 agcctgggac agaaccctac agaggccgag ctgcaggaca tgatcaacga ggttgacgct
     2401 gatggcgacg gaactatcga cttccccgag ttcctgacca tgatggccag aaagatgaag
     2461 tacagggaca cagaggaaga gatccgcgag gccttcggcg tgttcgataa ggatggaaac
     2521 ggctacatct ctgccgctga gctgagacac gtgatgacca acctgggcga gaagctgaca
     2581 gacgaagagg tggacgagat gatcagagag gccgacattg acggcgacgg gcaagtgaac
     2641 tacgaagagt ttgtccagat gatgaccgcc aagtgaaccg gtgtcgacga attctgcaga
     2701 tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca
     2761 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2821 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2881 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2941 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3001 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3061 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3421 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3481 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3541 ccaggcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3661 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3721 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3781 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3841 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3901 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3961 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4021 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4081 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4141 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4201 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4261 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4321 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4381 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4441 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4501 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4561 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4621 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4681 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4741 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4801 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4861 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4921 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4981 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5041 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5101 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5161 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5221 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5281 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5341 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5401 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5461 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5521 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5581 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     5641 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5701 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5761 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5821 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5881 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5941 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6001 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6061 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6121 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6181 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6241 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6301 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6361 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6421 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6481 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6541 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6601 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6661 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6721 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6781 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6841 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6901 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6961 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7021 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7081 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7141 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
