LOCUS       Exported                6340 bp DNA     circular SYN 16-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    mEos3.2-cortactin
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6340)
  AUTHORS   Shiqiong
  TITLE     Direct Submission
  JOURNAL   Exported Oct 13, 2022 from SnapGene 6.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6340
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(587..613)
                     /label=Vector.REV
     primer_bind     595..615
                     /label=Fragment.FOR
     regulatory      598..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             604..1281
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             604..606
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="M"
     CDS             604..606
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="M"
     primer_bind     607..627
                     /label=pr223
     primer_bind     complement(1270..1291)
                     /label=Fragment.REV
     primer_bind     1272..1306
                     /label=Vector.FOR
     misc_feature    1318..2958
                     /label=CortactinMouse
     primer_bind     complement(1397..1417)
                     /label=Cort_N_R
     polyA_signal    3119..3240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3156..3175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3210..3229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3247..3702)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3384..3405)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3596..3615
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3729..3833
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3835..4192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3858..3878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4043..4178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4105..4124
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4128..4139)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4227..5021
                     /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))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4281..4300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4891..4910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5209..5228)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5253..5300
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5629..6217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6118..6137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      601 accatgagtg cgattaagcc agacatgaag atcaaactcc gtatggaagg caacgtaaac
      661 gggcaccact ttgtgatcga cggagatggt acaggcaagc cttttgaggg aaaacagagt
      721 atggatcttg aagtcaaaga gggcggacct ctgccttttg cctttgatat cctgaccact
      781 gcattccatt acggcaacag ggtattcgcc aaatatccag acaacataca agactatttt
      841 aagcagtcgt ttcctaaggg gtattcgtgg gaacgaagct tgactttcga agacgggggc
      901 atttgcaacg ccagaaacga cataacaatg gaaggggaca ctttctataa taaagttcga
      961 ttttatggta ccaactttcc cgccaatggt ccagttatgc agaagaagac gctgaaatgg
     1021 gagccctcca ctgagaaaat gtatgtgcgt gatggagtgc tgacgggtga tattgagatg
     1081 gctttgttgc ttgaaggaaa tgcccattac cgatgtgact tcagaactac ttacaaagct
     1141 aaggagaagg gtgtcaagtt accaggcgcc cactttgtgg accactgcat tgagatttta
     1201 agccatgaca aagattacaa caaggttaag ctgtatgagc atgctgttgc tcattctgga
     1261 ttgcctgaca atgccagacg atccggactc agatctcgag ctcaagcttc gaattctatg
     1321 tggaaagcct ctgcaggcca tgctgtgtcc atcacgcagg atgatggagg agctgatgac
     1381 tgggagactg atcctgattt tgtgaatgat gtgagtgaaa aggagcagag atggggtgct
     1441 aaaaccgtgc agggatcggg gcaccaggaa cacatcaaca ttcacaagct tcgagagaat
     1501 gtcttccaag aacaccagac gctcaaggag aaggagctgg aaacgggacc caaggcttcc
     1561 cacggctatg gcgggaagtt cggtgtggag caggatagga tggacagatc agccgtgggc
     1621 catgagtacc agtcgaagct ttccaagcac tgctcacaag tggactcggt ccggggcttc
     1681 ggaggcaagt tcggtgtcca gatggacagg gtggatcagt ctgctgtagg ctttgaatac
     1741 caggggaaga ctgagaagca tgcctcccag aaagactact ctagtggctt cggtggcaaa
     1801 tacggtgtgc aagctgaccg tgtagacaag agtgccgtgg gctttgacta ccagggcaag
     1861 acggagaagc atgagtctca gaaagattac tccaaaggtt ttggtggcaa atatgggatt
     1921 gacaaggaca aggtggataa aagtgctgtg ggctttgagt atcaaggcaa gacagagaag
     1981 cacgaatccc agaaagacta tgtaaaaggc tttggaggaa agtttggtgt gcagacagac
     2041 agacaggaca agtgtgccct tggctgggac catcaggaga agctgcagct gcatgaatcc
     2101 caaaaagact ataagactgg tttcggaggc aaatttggtg ttcagtccga gaggcaggac
     2161 tcctccgctg tggggtttga ttacaaggag agattggcca agcacgagcc ccagcaagac
     2221 tatgccaaag gattcggcgg gaagtatggg gtgcagaagg atcggatgga caagaatgca
     2281 tccacctttg aagaagtggt ccaggtgcca tctgcctatc agaagactgt ccccattgag
     2341 gccgtaacca gcaaaaccag taatatccgt gctaactttg aaaacctggc aaaggagaga
     2401 gagcaggagg acaggcggaa ggcagaagcc gagagagctc agcggatggc caaagaaaga
     2461 caggagcagg aggaggcgcg caggaagctg gaagagcaag ccagagccaa gaagcagacg
     2521 ccccctgcat cccctagtcc tcaaccaatt gaagacagac caccctccag ccccatctat
     2581 gaggatgcag ctccgttcaa ggccgagccg agctaccgag gtagcgaacc tgagcctgag
     2641 tacagcatcg aggccgcagg cattcctgag gctggcagcc agcaaggcct gacctataca
     2701 tcagagcccg tgtacgagac tacagaggct cctggccact atcaagcaga ggatgacacc
     2761 tacgatgggt atgagagtga cctgggcatc acagccatcg ccctgtatga ctaccaggct
     2821 gctggcgatg atgagatctc ctttgaccct gatgacatca tcaccaacat agaaatgatt
     2881 gacgatggct ggtggcgtgg ggtgtgcaag ggcagatacg ggctcttccc agccaactat
     2941 gtggagctgc ggcagtagga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
     3001 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3061 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3241 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3301 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3361 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3421 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3481 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3541 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3601 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3661 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3841 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4021 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4141 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4201 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4261 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4321 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4381 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4441 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4501 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4561 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4621 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4681 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4741 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4801 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4861 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4921 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4981 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5041 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5101 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5161 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5221 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5281 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5341 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5401 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5461 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5521 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5581 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5641 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5701 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5761 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5821 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5881 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5941 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6001 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6061 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6301 ttctgtggat aaccgtatta ccgccatgca ttagttatta
//
