LOCUS       Exported                6446 bp DNA     circular SYN 29-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6446)
  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..6446
                     /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..596
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     mRNA            597..1379
                     /label=IL2
     misc_feature    1395..2378
                     /label=AH
     primer_bind     2397..2426
                     /label=Primer 2
     CDS             2403..3113
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3084..3113)
                     /label=Primer 1
     polyA_signal    3234..3355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3362..3817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3844..3948
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3950..4307
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4158..4293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4342..5136
                     /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    5368..5415
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      5744..6332
                     /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 gctagcatgg
      601 attcatacct gctgatgtgg ggactgctca cgttcatcat ggtgcctggc tgccaggcag
      661 agctctgtga cgatgacccg ccagagatcc cacacgccac attcaaagcc atggcctaca
      721 aggaaggaac catgttgaac tgtgaatgca agagaggttt ccgcagaata aaaagcgggt
      781 cactctatat gctctgtaca ggaaactcta gccactcgtc ctgggacaac caatgtcaat
      841 gcacaagctc tgccactcgg aacacaacga aacaagtgac acctcaacct gaagaacaga
      901 aagaaaggaa aaccacagaa atgcaaagtc caatgcagcc agtggaccaa gcgagccttc
      961 caggtcactg cagggaacct ccaccatggg aaaatgaagc cacagagaga atttatcatt
     1021 tcgtggtggg gcagatggtt tattatcagt gcgtccaggg atacagggct ctacacagag
     1081 gtcctgctga gagcgtctgc aaaatgaccc acgggaagac aaggtggacc cagccccagc
     1141 tcatatgcac aggtgaaatg gagaccagtc agtttccagg tgaagagaag cctcaggcaa
     1201 gccccgaagg ccgtcctgag agtgagactt cctgcctcgt cacaacaaca gattttcaaa
     1261 tacagacaga aatggctgca accatggaga cgtccatatt tacaacagag taccaggtag
     1321 cagtggccgg ctgtgttttc ctgctgatca gcgtcctcct cctgagtggg ctcacctgga
     1381 agggaaaatt ccaaagaact cgtgtccctc gagctgaatc tggtgatagc cttggttctg
     1441 aagatcgtga tcttctttac agcattgatg catatagatc tcaaagattc aaagaaacag
     1501 aacgtccatc aataaagcag gtgattgttc ggaaggaaga tgttacttca aaactggatg
     1561 aaaaaaataa tgcctttcct tgtcaagtta atatcaaaca gaaaatgcag gaactcaata
     1621 acgaaataaa tatgcaacag acagtgatct atcaagctag ccaggatctt aactgctgtg
     1681 ttgatgaaga acatggaaaa gggtccctag aagaagctaa agcagaaaga cttcttctaa
     1741 ttgcaactgg gaagagaaca cttttgattg atgaattgaa taaattgaag aacgaaggac
     1801 ctcagaggaa gaataaggct agtccccaaa gtgaatttat gccatccaaa ggatcagtta
     1861 ctttgtcaga aatccgcttg cctctaaaag cagattttgt ctgcagtacg gttcagaaac
     1921 cagatgcagc aaattactat tacttaatta tactaaaagc aggagctgaa aatatggtag
     1981 ccacaccatt agcaagtact tcaaactctc ttaacggtga tgctctgaca ttcactacta
     2041 catttactct gcaagatgta tccaatgact ttgaaataaa tattgaagtt tacagcttgg
     2101 tgcaaaagaa agatccctca ggccttgata agaagaaaaa aacatccaag tccaaggcta
     2161 ttactccaaa gcgactcctc acatctataa ccacaaaaag caacattcat tcttcagtca
     2221 tggccagtcc aggaggtctt agtgctgtgc gaaccagcaa cttcgccctt gttggatctt
     2281 acacattatc attgtcttca gtaggaaata ctaagtttgt tctggacaag gtcccctttt
     2341 tatcttcttt ggaaggtcat atttatttaa aaataaaatg tcaagtgcga ccggtcgcca
     2401 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2461 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2521 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     2581 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     2641 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2701 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2761 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2821 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2881 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2941 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     3001 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3061 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tagcggccgc
     3121 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3181 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3361 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3421 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3481 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3541 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3601 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3661 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3721 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3781 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3841 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3901 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3961 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4021 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4141 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4201 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4261 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4321 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4501 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4861 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5101 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5161 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5221 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5281 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5341 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5401 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5461 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5521 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5581 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5641 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5701 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5761 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5821 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5881 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5941 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6001 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6061 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6121 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6361 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6421 tggataaccg tattaccgcc atgcat
//
