LOCUS       Exported                6760 bp ds-DNA     circular SYN 11-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 11, 2025 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6760
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
     CDS             931..2955
                     /codon_start=1
                     /label=NES-FHA1-CKAR_FLIM
                     /translation="MLQNELALKLAGLDINKTGPVATMVSKGEELFTGVVPILVELDGD
                     VNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQ
                     HDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH
                     KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDN
                     HYLSTQSKLSKDPNEKRDHMVLLEFVTAARMHKFSQEQIGENIVCRVICTTGQIPIRDL
                     SADISQVLKEKRSIKKVWTFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTN
                     GTWLNGQKVEKNSNQLLSQGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDRSAGK
                     PGSGEGSTKGRFRRFQALKDKAKATGGSELMGSVQLADHYQQNTPIGDGPVLLPDNHYL
                     SYQSALSKDPNEKRDHMVLLERVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPIL
                     VELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTFGYGLMCFARY
                     PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKED
                     GNILGHKLEYNWNSVNVYIMADKQKNGIKVNFKIRHNIE"
     CDS             931..987
                     /codon_start=1
                     /label=NES from PKI alpha
                     /translation="MLQNELALKLAGLDINKTG"
     regulatory      994..1003
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1000..1683
                     /codon_start=1
                     /label=mEGFPdelta
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAA"
     misc_feature    1618..1620
                     /label=A206K (monomeric mutation)
     CDS             1693..2115
                     /codon_start=1
                     /label=FHA domain
                     /translation="KFSQEQIGENIVCRVICTTGQIPIRDLSADISQVLKEKRSIKKVW
                     TFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTNGTWLNGQKVEKNSNQLLS
                     QGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDR"
     primer_bind     2156..2196
                     /label=1092_H808ToH881_F
     primer_bind     complement(2156..2196)
                     /label=1093_H808ToH881_R
     CDS             2158..2199
                     /codon_start=1
                     /label=PKC substrate seq
                     /translation="RFRRFQALKDKAKA"
     misc_feature    2176..2178
                     /label=T2A mutation
     misc_feature    2185..2187
                     /label=D for FHA1
     misc_signal     2218..2952
                     /label=cp-sREACh @ E172, note no D173.
     misc_feature    2323..2325
                     /label=S208
     misc_feature    2368..2373
                     /label=R223,V224
     polyA_signal    3018..3152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3225..3582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3433..3568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    3433..3510
                     /label=SV40 ORI
     primer_bind     complement(3602..3618)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3831..4286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4618..4722
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4653..4681
                     /label=Amp prom
     CDS             4723..5583
                     /codon_start=1
                     /label=pRK5 Amp/Carb-R
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5732..6360
                     /direction=RIGHT
                     /label=ColE1 origin
     rep_origin      5754..6342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6666..6696
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        6666..6695
                     /label=lac
     misc_binding    6700..6722
                     /label=LacO
     protein_bind    6704..6720
                     /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)."
     primer_bind     6728..6748
                     /label=M13-rev
     primer_bind     6728..6744
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgccacc atgctgcaga acgagcttgc tcttaagttg
      961 gctggacttg atattaacaa gactggaccg gtcgccacca tggtgagcaa gggcgaggag
     1021 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1081 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1141 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1201 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1261 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1321 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1381 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1441 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1501 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1561 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     1621 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1681 gcccgcatgc ataagttttc tcaagaacag atcggcgaaa acattgtgtg cagggtcatt
     1741 tgtaccacgg gtcaaattcc catccgagat ttgtcagctg atatttcaca agtgcttaag
     1801 gaaaaacgat ccataaagaa agtttggaca tttggtagaa acccagcctg tgactatcat
     1861 ttaggaaaca tttcaagact gtcaaataag catttccaaa tactactagg agaagacggt
     1921 aaccttttat tgaatgacat ttccactaat gggacctggt taaatgggca aaaagtcgag
     1981 aagaacagca atcagttact gtctcaaggt gatgaaataa ccgttggtgt aggcgtggaa
     2041 tcagatattt tatctctggt cattttcata aacgacaaat ttaagcagtg cctcgagcag
     2101 aacaaagttg atcgctctgc aggtaagcca ggcagcggcg agggcagcac caagggccga
     2161 ttcagaagat tccaggcgtt gaaggataag gctaaggcca ccggcggcag cgagctcatg
     2221 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2281 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     2341 aagcgcgatc acatggtcct gctggagaga gtgaccgccg ccgggatcac tctcggcatg
     2401 gacgagctat acaagggtgg aactggaggt tcaatggtga gcaagggcga ggagctgttc
     2461 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2521 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctcatctgc
     2581 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2641 atgtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2701 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2761 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2821 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactg gaacagcgtc
     2881 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2941 cacaacatcg agtaagcggc cgcaattccc cggggatcct ctagagtcga cctgcagaag
     3001 cttggccgcc atggcccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3061 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3121 aactcatcaa tgtatcttat catgtctgga tcgatcggga attaattcgg cgcagcacca
     3181 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3241 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3301 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3361 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3421 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3481 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3541 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt aacagcttgg
     3601 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3661 gccttgcagc acatcccccc ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3721 gcccttccca acagttgcgt agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     3781 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3841 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3901 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3961 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4021 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4081 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4141 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     4201 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4261 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4321 cgcatagtta agccaactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4381 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4441 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4501 cgcgcgaggc agtattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4561 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4621 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4681 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4741 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4801 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4861 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4921 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgatga cgccgggcaa
     4981 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5041 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5101 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5161 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5221 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgccagcagc aatggcaaca
     5281 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5341 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5401 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5461 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5521 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5581 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5641 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5701 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5761 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5821 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5881 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     5941 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6001 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6061 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6121 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     6181 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6241 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6301 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6361 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6421 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6481 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6541 ccgcctctcc ccgcgcgttg gccgattcat taatccagct ggcacgacag gtttcccgac
     6601 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
     6661 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6721 tttcacacag gaaacagcta tgaccatgat tacgaattaa
//
