LOCUS       Exported                9880 bp ds-DNA     circular SYN 12-OCT-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9880)
  AUTHORS   Demo User
  TITLE     Direct Submission
  JOURNAL   Exported Aug 30, 2022 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9880
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      50..371
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     regulatory      377..698
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     5'UTR           799..929
                     /label=Translational enhancer
                     /note="translational enhancer 5'-UTR from tobacco etch 
                     virus"
     misc_feature    931..1606
                     /label=NC Frame 1
     misc_feature    1608..2129
                     /label=nEYFP
     regulatory      2151..2361
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     CDS             complement(2471..3124)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVVQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     regulatory      3223..3433
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     misc_feature    3443..4119
                     /label=NC Frame 2
     misc_feature    4135..4329
                     /label=CEYFP
     5'UTR           4338..4468
                     /label=Translational enhancer
                     /note="translational enhancer 5'-UTR from tobacco etch 
                     virus"
     regulatory      complement(4569..4890)
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     regulatory      4896..5217
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     misc_feature    5726..5750
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5841..6429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6530..7345)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7636..8071
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    8204..8226
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      8250..8501
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8541..9335)
                     /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(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(9369..9548)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 aagcttgttg taaaacgacg gccagtgccg gcgcgccacc ggtcaacatg tggagcacga
       61 cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga
      121 gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg
      181 tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga
      241 taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc
      301 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
      361 ttgatgtgat aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac
      421 agtctcagaa gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct
      481 cctcggattc cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg
      541 tggctcctac aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc
      601 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      661 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
      721 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
      781 ggagaggacg tcgagagttc tcaacacaac atatacaaaa caaacgaatc tcaagcaatc
      841 aagcattcta cttctattgc agcaatttaa atcatttctt ttaaagcaaa agcaattttc
      901 tgaaaatttt caccatttac gaacgatagc cagtggtctc tgtccagtcc tggcctcgtc
      961 ggccattctc gactaagttg gcagcatcac ccgacgcact ttgcgccgaa taaatacctg
     1021 tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat accgggaagc
     1081 cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc
     1141 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     1201 ctaaggaagc taaacttttg ctgacgagaa cagggactgg tgaaatgcag tttaaggttt
     1261 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     1321 acacgcctgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     1381 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     1441 ccgatatggc cagtgtgccg gtatccgtta tcggggaaga agtggctgat ctcagccacc
     1501 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     1561 cccttataca cagggccagt ctggccactt gtggtctgct gagacccatg gtgagcaagg
     1621 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1681 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1741 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1801 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1861 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1921 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1981 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2041 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2101 acttcaagat ccgccacaac atcgaggact gaagatccac ctgatctaga gtccgcaaaa
     2161 atcaccagtc tctctctaca aatctatctc tctctatttt tctccagaat aatgtgtgag
     2221 tagttcccag ataagggaat tagggttctt atagggtttc gctcatgtgt tgagcatata
     2281 agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc
     2341 ctaaaaccaa aatccagtga cgcttatggg accgacatat cagtatatat tcttataccg
     2401 caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc ggatccacgc
     2461 ggcgtttacg ccccccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     2521 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     2581 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     2641 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     2701 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     2761 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     2821 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2881 gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa
     2941 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     3001 taatatccag ctgaacggtc tggttatagg tacattgaac aactgactga aatgcctcaa
     3061 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     3121 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     3181 atcttatttc attatggtga aagttggaac ctcttaccgg gcgtcactgg attttggttt
     3241 taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat actaagggtt
     3301 tcttatatgc tcaacacatg agcgaaaccc tataagaacc ctaattccct tatctgggaa
     3361 ctactcacac attattctgg agaaaaatag agagagatag atttgtagag agagactggt
     3421 gatttttgcg gactctagat caccactgga aacatccagg tctggccaga ctggccctgt
     3481 gtataaggga gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
     3541 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggatacc ggcacactgg
     3601 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
     3661 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
     3721 caggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
     3781 tataggtgta aaccttaaac tgcatttcac cagtccctgt tctcgtcagc aaaagtttag
     3841 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     3901 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
     3961 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
     4021 cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttagtcg
     4081 agaatggccg acgaggccac gcatggtcga gccaactcta gatctgagtc cggacttgta
     4141 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     4201 atcgcgcttc tcgttggggt ctttgctcag ggcggactgg tagctcaggt agtggttgtc
     4261 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
     4321 cacgctgccg cccatggcta tcgttcgtaa atggtgaaaa ttttcagaaa attgcttttg
     4381 ctttaaaaga aatgatttaa attgctgcaa tagaagtaga atgcttgatt gcttgagatt
     4441 cgtttgtttt gtatatgttg tgttgagaac tctcgacgtc ctctccaaat gaaatgaact
     4501 tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc
     4561 agtggagata tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc
     4621 cacgatgctc ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttc
     4681 aacgatggcc tttcctttat cgcaatgatg gcatttgtag gagccacctt ccttttccac
     4741 tatcttcaca ataaagtgac agatagctgg gcaatggaat ccgaggaggt ttccggatat
     4801 taccctttgt tgaaaagtct caattgccct ttggtcttct gagactgtat ctttgatatt
     4861 tttggagtag acaagtgtgt cgtgctccac catgttatca catcaatcca cttgctttga
     4921 agacgtggtt ggaacgtctt ctttttccac gatgctcctc gtgggtgggg gtccatcttt
     4981 gggaccactg tcggcagagg catcttcaac gatggccttt cctttatcgc aatgatggca
     5041 tttgtaggag ccaccttcct tttccactat cttcacaata aagtgacaga tagctgggca
     5101 atggaatccg aggaggtttc cggatattac cctttgttga aaagtctcaa ttgccctttg
     5161 gtcttctgag actgtatctt tgatattttt ggagtagaca agtgtgtcgt gctccacatg
     5221 ttgaccggtg gcgcgccggc actggccgtc gttttacaac gaattcctgc agcccggggg
     5281 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     5341 gagggttaat tccgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5401 atccgctcac aattccacac aacatacgag ccggaaghca taaagtgtaa agcctggggt
     5461 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5521 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5581 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5641 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5701 aacgcaggaa agaacatgaa ggccttgaca ggatatattg gcgggtaaac taagtcgctg
     5761 tatgtgtttg tttgagatct catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6001 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6241 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6481 tcacgttaag ggattttggt catggttaca accaattaac caattctgat tagaaaaact
     6541 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     6601 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     6661 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     6721 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     6781 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     6841 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     6901 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     6961 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     7021 cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     7081 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     7141 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     7201 catcgggctt cccatacaat ccatagattg tcgcacctga ttgcccgaca ttatcgcgag
     7261 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag
     7321 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     7381 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     7441 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     7501 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     7561 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     7621 cgattcaggc gatccccatc caacagcccg ccgtcgagcg ggctttttta tccccggaag
     7681 cctgtggata gagggtagtt atccacgtga aaccgctaat gccccgcaaa gccttgattc
     7741 acggggcttt ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg
     7801 aaatcgctaa tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa
     7861 aggcacgtga gaacgctaat agccctttca gatcaacagc ttgcaaacac ccctcgctcc
     7921 ggcaagtagt tacagcaagt agtatgttca attagctttt caattatgaa tatatatatc
     7981 aattattggt cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga
     8041 caaccgcaag cggttgccca ccgtcgagcg ccagcgcctt tgcccacaac ccggcggccg
     8101 gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc
     8161 aacctctcgg gcttctggat ttccgatccc cggaattaga tcttggcagg atatattgtg
     8221 gtgtaacgtt atcagcttgc atgccggtcg atctagtaac atagatgaca ccgcgcgcga
     8281 taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg
     8341 cgggactcta atcaaaaaac ccatctcata aataacgtca tgcattacat gttaattatt
     8401 acatgcttaa cgtaattcaa cagaaattat atgataatca tcgcaagacc ggcaacagga
     8461 ttcaatctta agaaacttta ttgccaaatg tttgaacgat ctgcttgact ctagctagag
     8521 tccgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     8581 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     8641 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     8701 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     8761 caggcatcgc cctgggtcac gacgagatcc tcgccgtcgg gcatccgcgc cttgagcctg
     8821 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     8881 agaccggctt ccatccgagt acgtcctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     8941 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     9001 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     9061 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     9121 gtggccagcc acgatagccg cgctgcctcg tcttggagtt cattcagggc accggacagg
     9181 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     9241 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     9301 ggagaacctg cgtgcaatcc atcttgttca atcatgcctc gatcgagttg agagtgaata
     9361 tgagactcta attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa
     9421 gaaatatttg ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct
     9481 gacgtatgtg cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt
     9541 tgcggttctg tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata
     9601 acgtgactcc cttaattctc atgtatcgat aacattaacg tttacaattt cgcgccattc
     9661 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9721 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9781 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
     9841 attgggtacc gggccccccc tcgaggtcga cggtatcgat
//
