LOCUS       Exported                7676 bp ds-DNA    circular SYN 22-JAN-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1_rsAKARev
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7676)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported dinsdag 26 jan 2021 from SnapGene 2.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7676
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             914..1621
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /note="mTFP0.7"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTNAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTLKWEPSTEILYVRDGVLV
                     GDIKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARYSTGGMDELYK"
     CDS             1634..2056
                     /codon_start=1
                     /note="FHA1"
                     /translation="KFSQEQIGENIVCRVICTTGQIPIRDLSADISQVLKEKRSIKKVW
                     TFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTNGTWLNGQKVEKNSNQLLS
                     QGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDR"
     CDS             2063..2410
                     /codon_start=1
                     /note="EV-linker"
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             2417..2440
                     /codon_start=1
                     /note="PKA substrate"
                     /translation="LRRATLVD"
     CDS             2456..3190
                     /codon_start=1
                     /note="cpVenus[172]"
                     /translation="MGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     polyA_signal    3276..3500
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3546..3974
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3988..4317
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      4168..4303
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             4384..5178
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5352..5473
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5522..5538)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5546..5562
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     promoter        complement(5570..5600)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    5615..5636
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5924..6509)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6680..7540)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7541..7645)
                     /gene="bla"
                     /note="AmpR promoter"
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 gcttggtacc
      901 gagctcggat cccatggtga gcaagggcga ggagaccaca atgggcgtaa tcaagcccga
      961 catgaagatc aagctgaaga tggagggcaa cgtgaatggc cacgccttcg tgatcgaggg
     1021 cgagggcgag ggcaagccct acgacggcac caacaccatc aacctggagg tgaaggaggg
     1081 agcccccctg cccttctcct acgacattct gaccaacgcc ttcgcttacg gcaacagggc
     1141 cttcaccaag taccccgacg acatccccaa ctacttcaag cagtccttcc ccgagggcta
     1201 ctcttgggag cgcaccatga ccttcgagga caagggcatc gtgaaggtga agtccgacat
     1261 ctccatggag gaggactcct tcatctacga gatacacctc aagggcgaga acttcccccc
     1321 caacggcccc gtgatgcaga agaagaccct gaagtgggag ccctccaccg agatcctgta
     1381 cgtgcgcgac ggcgtgctgg tgggcgacat caagcacaag ctgctgctgg agggcggcgg
     1441 ccaccaccgc gttgacttca agaccatcta cagggccaag aaggcggtga agctgcccga
     1501 ctaccacttt gtggaccacc gcatcgagat cctgaaccac gacaaggact acaacaaggt
     1561 gaccgtttac gagagcgccg tggcccgcta ctccaccggc ggcatggacg agctgtacaa
     1621 gagatctatg cataagtttt ctcaagaaca gatcggcgaa aacattgtgt gcagggtcat
     1681 ttgtaccacg ggtcaaattc ccatccgaga tttgtcagct gatatttcac aagtgcttaa
     1741 ggaaaaacga tccataaaga aagtttggac atttggtaga aacccagcct gtgactatca
     1801 tttaggaaac atttcaagac tgtcaaataa gcatttccaa atactactag gagaagacgg
     1861 taacctttta ttgaatgaca tttccactaa tgggacctgg ttaaatgggc aaaaagtcga
     1921 gaagaacagc aatcagttac tgtcccaagg tgatgaaata accgttggtg taggcgtgga
     1981 atcagatatt ttatctctgg tcattttcat aaacgacaaa tttaagcagt gcctggagca
     2041 gaacaaagtt gatcgcggta ccagtgctgg tggtagtgct ggtggtagtg ctggtggtag
     2101 tgctggtggt agtgctggtg gttccggcag tgctggtggt agtgctggtg gtagtaccag
     2161 tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggttc
     2221 cggcagtgct ggtggtagtg ctggtggtag taccagtgct ggtggtagtg ctggtggtag
     2281 tgctggtggt agtgctggtg gtagtgctgg tggttccggc agtgctggtg gtagtgctgg
     2341 tggtagtacc agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggtag
     2401 tgctggtggt tccggattga ggcgcgcgac gctggttgac ggcggcagcg agctcatggg
     2461 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2521 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
     2581 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2641 cgagctgtac aagggaggta ccggtggatc tatggtgagc aagggcgagg agctgttcac
     2701 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2761 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     2821 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggccttca
     2881 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2941 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3001 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3061 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3121 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     3181 caacatcgag taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc
     3241 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3301 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3361 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3421 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3481 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3541 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3961 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4081 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4141 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4201 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4261 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4321 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4381 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4441 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4501 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4561 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4621 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4681 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4741 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4801 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4861 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4921 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4981 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5041 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5101 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5161 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5221 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5281 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5341 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5401 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5461 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5521 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5581 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5641 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5701 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5761 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5821 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5881 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5941 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6001 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6061 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6121 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6181 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6241 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6301 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6361 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6421 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     6481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6661 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6721 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6781 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6841 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6901 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6961 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7021 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7081 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7141 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7201 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7261 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7321 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7381 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7441 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7501 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7561 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7621 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
