LOCUS       Exported                6959 bp DNA     circular SYN 18-OCT-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6959)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Jun 7, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    591..595
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     mRNA            596..1378
                     /product="interleukin-2 receptor alpha-subunit"
     misc_feature    1394..2239
                     /label=AH deltaC2
                     /note="Anillin homology domain"
     CDS             2258..2311
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2330..3037
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2330..2353
                     /label=Primer 2
     primer_bind     complement(3011..3037)
                     /label=Primer 1
     CDS             3044..3625
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase RhoA"
                     /label=RhoA (Q63L)
                     /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD
                     IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK
                     HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT
                     KDGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    3747..3868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3875..4330)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4357..4461
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4463..4820
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4671..4806
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4855..5649
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    5881..5928
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      6257..6845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagatgga
      601 ttcatacctg ctgatgtggg gactgctcac gttcatcatg gtgcctggct gccaggcaga
      661 gctctgtgac gatgacccgc cagagatccc acacgccaca ttcaaagcca tggcctacaa
      721 ggaaggaacc atgttgaact gtgaatgcaa gagaggtttc cgcagaataa aaagcgggtc
      781 actctatatg ctctgtacag gaaactctag ccactcgtcc tgggacaacc aatgtcaatg
      841 cacaagctct gccactcgga acacaacgaa acaagtgaca cctcaacctg aagaacagaa
      901 agaaaggaaa accacagaaa tgcaaagtcc aatgcagcca gtggaccaag cgagccttcc
      961 aggtcactgc agggaacctc caccatggga aaatgaagcc acagagagaa tttatcattt
     1021 cgtggtgggg cagatggttt attatcagtg cgtccaggga tacagggctc tacacagagg
     1081 tcctgctgag agcgtctgca aaatgaccca cgggaagaca aggtggaccc agccccagct
     1141 catatgcaca ggtgaaatgg agaccagtca gtttccaggt gaagagaagc ctcaggcaag
     1201 ccccgaaggc cgtcctgaga gtgagacttc ctgcctcgtc acaacaacag attttcaaat
     1261 acagacagaa atggctgcaa ccatggagac gtccatattt acaacagagt accaggtagc
     1321 agtggccggc tgtgttttcc tgctgatcag cgtcctcctc ctgagtgggc tcacctggaa
     1381 gggaaaattc caaagaactc gtgtccctcg agctgaatct ggtgatagcc ttggttctga
     1441 agatcgtgat cttctttaca gcattgatgc atatagatct caaagattca aagaaacaga
     1501 acgtccatca ataaagcagg tgattgttcg gaaggaagat gttacttcaa aactggatga
     1561 aaaaaataat gcctttcctt gtcaagttaa tatcaaacag aaaatgcagg aactcaataa
     1621 cgaaataaat atgcaacaga cagtgatcta tcaagctagc caggctctta actgctgtgt
     1681 tgatgaagaa catggaaaag ggtccctaga agaagctgaa gcagaaagac ttcttctaat
     1741 tgcaactggg aagagaacac ttttgattga tgaattgaat aaattgaaga acgaaggacc
     1801 tcagaggaag aataaggcta gtccccaaag tgaatttatg ccatccaaag gatcagttac
     1861 tttgtcagaa atccgcttgc ctctaaaagc agattttgtc tgcagtacgg ttcagaaacc
     1921 agatgcagca aattactatt acttaattat actaaaagca ggagctgaaa atatggtagc
     1981 cacaccatta gcaagtactt caaactctct taacggtgat gctctgacat tcactactac
     2041 atttactctg caagatgtat ccaatgactt tgaaataaat attgaagttt acagcttggt
     2101 gcaagctgtg cgaaccagca acttcgccct tgttggatct tacacattat cattgtcttc
     2161 agtaggaaat actaagtttg ttctggacaa ggtccccttt ttatcttctt tggaaggtca
     2221 tatttattta aaaataaaat gtcaagtgcg accggtcgag ggcaggggaa gcctgctcac
     2281 ctgcggcgac gtggaggaaa accctggccc cgccccagga tccaagctta tggtgagcaa
     2341 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     2401 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2461 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     2521 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     2581 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2641 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     2701 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     2761 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     2821 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     2881 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     2941 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     3001 ccgccactcc accggcggca tggacgagct gtacaaggaa ttcatggctg ccatccggaa
     3061 gaaactggtg attgttggtg atggagcctg tggaaagaca tgcttgctca tagtcttcag
     3121 caaggaccag ttcccagagg tgtatgtgcc cacagtgttt gagaactatg tggcagatat
     3181 cgaggtggat ggaaagcagg tagagttggc tttgtgggac acagctgggc tggaagatta
     3241 tgatcgcctg aggcccctct cctacccaga taccgatgtt atactgatgt gtttttccat
     3301 cgacagccct gatagtttag aaaacatccc agaaaagtgg accccagaag tcaagcattt
     3361 ctgtcccaac gtgcccatca tcctggttgg gaataagaag gatcttcgga atgatgagca
     3421 cacaaggcgg gagctagcca agatgaagca ggagccggtg aaacctgaag aaggcagaga
     3481 tatggcaaac aggattggcg cttttgggta catggagtgt tcagcaaaga ccaaagatgg
     3541 agtgagagag gtttttgaaa tggctacgag agctgctctg caagctagac gtgggaagaa
     3601 aaaatctggt tgccttgtct tgtgagcggc cgcgactcta gatcataatc agccatacca
     3661 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3721 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3781 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3841 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3901 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3961 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4021 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4081 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4141 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4201 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4261 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4321 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4381 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4441 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4501 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4621 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4681 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4741 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4801 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4861 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4921 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4981 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5041 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5101 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5161 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5221 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5281 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5341 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5401 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5461 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5521 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5581 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5641 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5701 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5761 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5821 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5881 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5941 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6001 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6061 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6901 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
