LOCUS       Exported                6172 bp ds-DNA     circular SYN 20-NOV-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6172)
  AUTHORS   L Preston
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6172
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        69..309
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        565..776
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        787..805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      821..830
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             836..856
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             2093..2140
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /note="color: #993366"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2147..2170
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /note="color: #cc99b2"
                     /translation="DYKDDDDK"
     CDS             2186..2239
                     /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."
                     /note="color: #cc99b2
                     Cleavage site after base 2236"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2240..2947
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /note="color: #05fd14"
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      2492..2501
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     polyA_signal    3205..3339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(3548..4136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(4307..5167)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4307 .. 5098 / #ccffcc
                        2: 5099 .. 5167 / #ccffcc / signal sequence
                     Cleavage site after base 5098"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5168..5272)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(5554..6009)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     repeat_region   join(6084..6172,1..52)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
ORIGIN
        1 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
       61 ttaattaaga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
      121 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
      181 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
      241 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
      301 ggaaaggacg aaacaccgtg gtggctgcgg gaatctcaga caccttaaac gctcatggag
      361 gctatgaaaa tggtctgcga agtgagaatc acgcgacttt agtcgtgtga ggttcaaatg
      421 tcttcctggg acgaagacaa ggccggcatg gtcccagcct cctcgctggc gccggctggg
      481 caacatgctt cggcatggcg aatgggactt tttttacgcg ttaggtcttg aaaggagtgg
      541 gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      601 tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg
      661 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      721 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacg
      781 ggaaattaat acgactcact atagggaccg gtgcggccgc gccaccatgg gatctccaaa
      841 aaagaaacgg aaggtgggat ctatgatccg gaacaaggcc ttcgtggtga ggctgtaccc
      901 caacgccgcc cagacagagc tgatcaatag aaccctgggc agcgccaggt tcgtgtacaa
      961 tcacttcctg gccaggagga tcgccgccta caaggagtcc ggcaagggcc tgacatacgg
     1021 ccagacatcc agcgagctga ccctgctgaa gcaggccgag gagacatcct ggctgagcga
     1081 ggtggacaag tttgccctgc agaacagcct gaagaatctg gagaccgcct acaagaactt
     1141 cttcagaaca gtgaagcaga gcggcaagaa ggtgggcttt cccaggttca gaaagaagag
     1201 aacaggcgag agctacagga cacagtttac aaataacaac atccagatcg gcgagggcag
     1261 gctgaagctg cctaagctgg gctgggttaa gacaaagggc cagcaggata ttcagggcaa
     1321 gatcctgaat gtgacagtga gaaggattca tgagggccac tacgaggcca gcgtgctgtg
     1381 tgaggtggag atcccctacc tgcccgccgc tcctaagttc gccgccggag ttgatgtggg
     1441 catcaaggac tttgccatcg tgaccgacgg cgtgagattt aagcacgagc agaaccctaa
     1501 gtactacagg agcacactga agaggctgag gaaggcccag cagacactgt ccagaagaaa
     1561 gaagggcagc gcccggtacg gcaaggccaa gacaaagctg gccaggattc ataagagaat
     1621 cgtgaataag aggcaggact ttctgcacaa gctgaccaca agcctggtga gggagtacga
     1681 gatcatcggc acagagcacc tgaagcctga taacatgagg aagaacagga gactggccct
     1741 gtccatcagc gatgccggct ggggagagtt tatcagacag ctggagtaca aggccgcctg
     1801 gtacggcaga ctggtgagca aggtgagccc ttacttcccc agctcccagc tgtgtcacga
     1861 ctgcggcttc aagaatcccg aggtgaagaa cctggccgtg agaacctgga cctgtcccaa
     1921 ctgcggcgag acccacgaca gagatgaaaa tgccgccctg aacatcagga gggaggccct
     1981 ggtggccgcc ggaattagcg acacactgaa tgcccacggc ggctacgtga gacctgcctc
     2041 tgccggaaat ggcctgaggt ccgagaacca cgccaccctg gtggtgggtt ccaaaaggcc
     2101 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatctgatt acaaagacga
     2161 tgacgataag gaattcggta gtggggaggg cagaggaagt ctgctaacat gcggtgacgt
     2221 cgaggagaat cctggcccag tgagcaaggg cgaggaggat aacatggcct ctctcccagc
     2281 gacacatgag ttacacatct ttggctccat caacggtgtg gactttgaca tggtgggtca
     2341 gggcaccggc aatccaaatg atggttatga ggagttaaac ctgaagtcca ccaagggtga
     2401 cctccagttc tccccctgga ttctggtccc tcatatcggg tatggcttcc atcagtacct
     2461 gccctaccct gacgggatgt cgcctttcca ggccgccatg gtagatggct ccggatacca
     2521 agtccatcgc acaatgcagt ttgaagatgg tgcctccctt actgttaact accgctacac
     2581 ctacgaggga agccacatca aaggagaggc ccaggtgaag gggactggtt tccctgctga
     2641 cggtcctgtg atgaccaact cgctgaccgc tgcggactgg tgcaggtcga agaaaactta
     2701 ccccaacgac aaaaccatca tcagtacctt taagtggagt tacaccactg gaaatggcaa
     2761 gcgctaccgg agcactgcgc ggaccaccta cacctttgcc aagccaatgg cggctaacta
     2821 tctgaagaac cagccgatgt acgtgttccg taagacggag ctcaagcact ccaagaccga
     2881 gctcaacttc aaggagtggc aaaaggcctt taccgatgtg atgggcatgg acgagctgta
     2941 caagtgaaag cttgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3001 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3061 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     3121 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3181 gggcactgac aattccgtgg tgttaacttg tttattgcag cttataatgg ttacaaataa
     3241 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3301 ttgtccaaac tcatcaatgt atcttatcat gtctggatcg gcgcgccagc ggccgcagga
     3361 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3421 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3481 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3541 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3601 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3661 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3721 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3781 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3841 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3901 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3961 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4021 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4081 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4141 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4201 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4261 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4321 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4381 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4441 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4501 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4561 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4621 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4681 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4741 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4801 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4861 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4921 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4981 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5041 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5101 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5161 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5221 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5281 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5341 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     5401 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5461 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     5521 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     5581 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     5641 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     5701 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5761 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     5821 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5881 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
     5941 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6001 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
     6061 gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc
     6121 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tg
//
