LOCUS       Exported                6628 bp ds-DNA     circular SYN 10-MAY-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 10, 2022 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6628
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli 
                     rrnBgene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli 
                     rrnBgene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(603..727)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    774..1707
                     /label=X3 5' flk region
     misc_feature    complement(1721..1889)
                     /label=DmTub56Dterm
     CDS             complement(1890..2609)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(2610..3986)
                     /label=AePUb promoter
     misc_feature    3987..4861
                     /label=X3 3' flk region
     misc_feature    4552..4842
                     /label=deleted in KI plasmid
     promoter        complement(4870..4888)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4893..4909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5022..5831
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5978..6566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttggtac cgggcccccc gctagcgtcg acggtatcga taagcttgat cggatccgga
      781 gagtacgttc gcattgccga agtaaaaact accaagaaaa ttcaacacag cggtggcgtt
      841 tactattcaa acgatgtggt aaaacgtaat ttagccacag tcctctcggg aaatctcaac
      901 gaagaaaccg aatatgtatc cttgaatgaa ttgccgtgca atatgcgatg cgagagcaat
      961 gttctctcgc aaaacgacac ccctcctggc ggaagcggaa aactgaatac ttcgggtacg
     1021 ggaggagaag caccctcggc aggagaaaac aacgaacatg ctataaaaag gggtgcacgc
     1081 gtgaaactgg atgcgcacgg taaggttatc tacagttcgg atagtctgaa gaggagaaag
     1141 ggtgcgcaca cgacctttgc gccgggtcca tttgtgaaag acgtaaatac ggcacagcca
     1201 ctgacgacca acactcctac tacaaacact acaactcggt cagcatcacc agcagcatcc
     1261 acagaaaatg gcaaatgtgc tgttgttgca tctccattac taagtggtgg tctagctaac
     1321 aggaaaccaa tagcagttaa accagtgata tcgcagtccg cattgaaaac gaaagcaatc
     1381 gtcaacacca atagtaatat aagtaatcat agcgacggag tggcagccca tcgtatggcc
     1441 ccaattgcca gtattcttcc caacaacagt acggtgcaag aagtggaaaa aatagctgca
     1501 gcatcacaca tcatgtcctc cggagttcta aaaggtgcat acgttaatgt acaggaatct
     1561 aaaccaccgt cttcaaaacc tcaatcgatt cactatatgg aagcaacaaa cttacagcat
     1621 ccccaaacag gtaagcaagc taaatcgcaa aagaataaca aaatagttgg tgcttcgcca
     1681 aaggctagga acgtaatatt tccctccggg gttcctatac gaaaccccaa caaaaaccat
     1741 aattgtttag acttgtgaac aaaattggat ccgactttat tgattacgtt gttaagagaa
     1801 caaatctttt acaactgaat tcatttgttc tcgtttcatt ttttttcgca aaacattgat
     1861 cgagaattcg attgatttcc gattcgaatt tacttgtaca gctcgtccat gccgagagtg
     1921 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     1981 ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     2041 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     2101 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
     2161 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     2221 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     2281 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     2341 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     2401 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     2461 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     2521 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2581 gtgaacagct cctcgccctt gctcaccatg gttgaaatct ctgtttagca gaaaaagaaa
     2641 cgaggaaacg cttcagtaat tggttgtgaa atgcaaactc tcatttgata ttgattcatt
     2701 gcctttggct tcgagcacga cacgacaggt tttaaacttg ttttgctttg tctgcgtttg
     2761 cagtcgcagg ccaagtgaaa aatatacact tgaaggtgat gacgtcacaa caacgcccta
     2821 ctttttagtg aaaaattaac ttgttttcga ctttgaacta cgtagtttgg aaattgcgta
     2881 tcttcaagtt ttcacgattt cctcaaggtt tttctcgata tgtgttaata ttaccttaat
     2941 gggtaattat caccacaata tttaatttta gaaatatggc gaattcgcgc ttgctctaga
     3001 aatttaattc aattagcagc gatgattcaa cgaaatatga ttatcgccgt gaatcacaat
     3061 tgggttttat caatgatgat gaaactgcgt tgcaaatttt cactaatcac tcaaagctca
     3121 atagtcgcca tcttggaaaa tagtttgctc attcaaagac aaaggaatca ttcaccaaac
     3181 tagttttcgc tcatagctat aatttcatca tttaatttac ctaccttcac tagaagattc
     3241 ccttcaccga aatgcacttt cacaaattta atagtaattg tcctttgaac aaagctttgt
     3301 tcactctgaa attttctcct ctggctaatt ggatcactct ttttcactag agacttcact
     3361 tcacttgcac tggcactgct tacttggacc gcgtaatgtt cactccacta ggaaacgtat
     3421 tcgattgagc tggttttgcc tttgcagggg cgttttatag acactgccgt agtggtggtt
     3481 gtacttctag aaaatttctg caatccattc atacatatac ctctgttcgg ttggatggct
     3541 ctaaatcgat cgactatggg taccatccct gtgtctgaat ggacagcaaa acgtgcttgt
     3601 gtctgttagt cgttcattct gtaccttgaa cgatgcagtt caacttctgg caaagacgtc
     3661 aatgtaccta ccattcgtgt atatggcata cagagaaaga tgtgcgaatg cctttttcga
     3721 tagagaaagg atttcctgat ttgatcgaca atttcggtag ctattcttag cttcgaatgt
     3781 aatctgataa ccaaatccag agacaaagca gtaatttggg aatttcactt gaatcattct
     3841 aattgtatca tttgctgtat tagtgcagtc agcaaagtga cgtcaaccct tctaaatcga
     3901 tatacttctg ggaagctttc tttcttgtct ggctcagctg gtgccaaggc aaattataat
     3961 tggattcaat gcacaagcta catgtaaaga cgcatgatac tcatcacatt agacgtaata
     4021 aggacattga cccagatagc aattctacta tcaagcgcag gaatagctat agaaatgcga
     4081 actcgttcaa aatcgatact gatgctgcga gttttaccga tttcttggaa gaacatgtaa
     4141 ataaacaaga ggaacggcta cagtcctccg cggtgaagaa tgatagcaat tcatataaca
     4201 agtataccca tttgcatgat caaaatagcg ataaggacga tggctacgaa gaaatcagcg
     4261 atattgactt ggacgaggat agcaaatcta ttgctcgttc gtttgaaaga ttactcaatc
     4321 tatcagatga ggtgtttggg cttcagaaga tggatagcac tagacccagt atcctggaga
     4381 agtgcaattt cgatgaatta gataccagta cagtggagaa taatgacgac gagttggaga
     4441 ttgtattacg tgacctgata ccttccacac ctaccggttg tagaagaagt ttctctattc
     4501 ctcttacacc cagtaagttg actatacctt cagatggggg tcagcatcgt gctagagttc
     4561 ttagcttcag cgctaccagc aacgacaact ctacagatat ctggtgagtc ggacgtttta
     4621 atataatttt caatggtagt gtttaacact catttctccc aataaatcac aaaagtacaa
     4681 attattgtca tctagacaca tttttaaata atcgccttta tcttggctct attctattct
     4741 aattattttt gttaaataag acaaatacac catgctttat aatgaaggcc attaatcaaa
     4801 aagttacaaa atgacaacca ttcaagtgca tgtccgactg tggcatagcg ccgtgtggac
     4861 ggcttatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     4921 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     4981 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     5041 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     5101 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     5161 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     5221 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     5281 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     5341 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     5401 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     5461 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     5521 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     5581 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     5641 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     5701 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     5761 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     5821 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     5881 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     5941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6121 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6601 ccttttgctg gccttttgct cacatgtt
//
