LOCUS       Exported                7097 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 7097)
  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..7097
                     /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..2377
                     /label=AH
                     /note="Anillin homology domain"
     CDS             2396..2449
                     /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             2468..3175
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2468..2491
                     /label=Primer 2
     primer_bind     complement(3149..3175)
                     /label=Primer 1
     CDS             3182..3763
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase RhoA"
                     /label=RhoA (Q63L)
                     /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD
                     IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK
                     HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT
                     KDGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    3885..4006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4013..4468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4601..4958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4809..4944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4993..5787
                     /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    6019..6066
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      6395..6983
                     /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 caggatctta actgctgtgt
     1681 tgatgaagaa catggaaaag ggtccctaga agaagctaaa 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 gcaaaagaaa gatccctcag gccttgataa gaagaaaaaa acatccaagt ccaaggctat
     2161 tactccaaag cgactcctca catctataac cacaaaaagc aacattcatt cttcagtcat
     2221 ggccagtcca ggaggtctta gtgctgtgcg aaccagcaac ttcgcccttg ttggatctta
     2281 cacattatca ttgtcttcag taggaaatac taagtttgtt ctggacaagg tccccttttt
     2341 atcttctttg gaaggtcata tttatttaaa aataaaatgt caagtgcgac cggtcgaggg
     2401 caggggaagc ctgctcacct gcggcgacgt ggaggaaaac cctggccccg ccccaggatc
     2461 caagcttatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2521 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2581 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2641 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2701 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2761 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2821 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2881 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2941 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     3001 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     3061 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3121 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggaatt
     3181 catggctgcc atccggaaga aactggtgat tgttggtgat ggagcctgtg gaaagacatg
     3241 cttgctcata gtcttcagca aggaccagtt cccagaggtg tatgtgccca cagtgtttga
     3301 gaactatgtg gcagatatcg aggtggatgg aaagcaggta gagttggctt tgtgggacac
     3361 agctgggctg gaagattatg atcgcctgag gcccctctcc tacccagata ccgatgttat
     3421 actgatgtgt ttttccatcg acagccctga tagtttagaa aacatcccag aaaagtggac
     3481 cccagaagtc aagcatttct gtcccaacgt gcccatcatc ctggttggga ataagaagga
     3541 tcttcggaat gatgagcaca caaggcggga gctagccaag atgaagcagg agccggtgaa
     3601 acctgaagaa ggcagagata tggcaaacag gattggcgct tttgggtaca tggagtgttc
     3661 agcaaagacc aaagatggag tgagagaggt ttttgaaatg gctacgagag ctgctctgca
     3721 agctagacgt gggaagaaaa aatctggttg ccttgtcttg tgagcggccg cgactctaga
     3781 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3841 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4021 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4081 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4141 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4201 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4261 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4321 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4381 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4441 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4561 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4621 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4681 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4801 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4861 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4921 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5161 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5521 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5941 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6061 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6121 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6181 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6541 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7081 gtattaccgc catgcat
//
