LOCUS       Exported                7721 bp ds-DNA     circular SYN 16-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7721)
  AUTHORS   Kwan KM, Fujimoto E, Grabher C, Mangum BD, Hardy ME, Campbell DS,
  TITLE     The Tol2kit: a multisite gateway-based construction kit for Tol2
  JOURNAL   Developmental Dynamics 236(11):3088-99 (2007).
  PUBMED    17937395
REFERENCE   2  (bases 1 to 7721)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 16, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     395.pDestTol2CG2, Tol2kit v1.0
FEATURES             Location/Qualifiers
     source          1..7721
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..22
                     /label=M13R
     primer_bind     1..22
                     /label=M13R(1)
     misc_recomb     37..161
                     /label=attR4
     misc_feature    complement(201..506)
                     /label=ccdB
                     /label=nonstandard type:  Resistance
     CDS             complement(848..1507)
                     /codon_start=1
                     /label=cmR
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_recomb     1616..1740
                     /label=attR3
     primer_bind     complement(1749..1764)
                     /label=M13F
     polyA_signal    1780..1977
                     /label=SV40 late polyA (from pCS2+)
     promoter        2047..2666
                     /label=crybb
     promoter        2718..2874
                     /label=fragment of minimal CMV promoter
     CDS             2875..3595
                     /codon_start=1
                     /label=codon optimized TagBFP
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN*SGPSR"
     polyA_signal    3627..3759
                     /label=SV40 late polyA (from pCS2+)
     misc_structure  complement(3766..4100)
                     /label=Tol2 exon1 bp1-335 of 461
     LTR             complement(4264..4282)
                     /label=Tol2 5' TIR
     promoter        complement(4368..4387)
                     /label=T3
                     /label=T3 promoter
     primer_bind     complement(4403..4424)
                     /label=M13R
     primer_bind     complement(4403..4424)
                     /label=M13R(1)
     rep_origin      complement(4751..5424)
                     /direction=LEFT
                     /label=pUC ori
     CDS             complement(5572..6429)
                     /codon_start=1
                     /label=ampR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     LTR             complement(7175..7191)
                     /label=Tol2 3' TIR
     misc_structure  complement(7364..7710)
                     /label=Tol2 exon4 136-482
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaac
       61 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
      121 ctacataata ctgtaaaaca caacatatcc agtcactatg gtcgacctgc agactggctg
      181 tgtataaggg agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg
      241 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg
      301 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt
      361 tcacggggga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc
      421 ccgggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt
      481 ttataggtgt aaaccttaaa ctgcatttca ccagcccctg ttctcgtcgg caaaagagcc
      541 gttcatttca ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg
      601 ttcggcacgc agacgacggg cttcattctg catggttgtg cttaccgaac cggagatatt
      661 gacatcatat atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc
      721 tgcttcatag catacctctt tttgacatac ttcgggtata catatcagta tatattctta
      781 taccgcaaaa atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc
      841 ctctagatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
      901 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
      961 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     1021 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     1081 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     1141 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     1201 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     1261 ccagctcacc gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa
     1321 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     1381 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     1441 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     1501 tctccatttt agcttcctta gctcctgaaa atctcgacgg atcctaactc aaaatccaca
     1561 cattatacga gccggaagca taaagtgtaa agcctggggg tgcctaatgc ggccgccata
     1621 gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat
     1681 ttaatatatt gatatttata tcattttacg tttctcgttc aactttatta tacatagttg
     1741 ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat gataagatac
     1801 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1861 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1921 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1981 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca gatcttcgag
     2041 caattggtgc tatcatcatg acaaagagaa aagaaatcag ttattagaaa tgagttatta
     2101 aaactattat gattagaaat atgtttcttt ctcttaaaca ggatttaagg ggaaaatata
     2161 cagggagcga aaaattttga cttcatctgt atatacatat aataaataat acatcaataa
     2221 attccatcta aatacgctac catatatata tatatatgta tatacacaca cacacacaca
     2281 ccatctaata tatgtcatgc tacattatta acttcagtat gaaatctact ctgctctgga
     2341 gtgtatcagc gagcgactcc tctgctcgga ttatcatcac gtgacctctg acctgaacac
     2401 agacctctct tagccatagg tatacagcct atagcataga gctcgggctg agagggaaag
     2461 tgccgccgtc agggctttcc attgacagta aaatgcttac ggccttctgg gcgatgcgat
     2521 gaccctgcgc ttcggcacgg ctgaactctg atgacggcgt gtttctattg tccgccgcgc
     2581 gccgctgtgc tgcccttgcc agcgtataaa agctgcggca gtggtgagga aggccagagc
     2641 gagccagcct tcccagcaca gctgtagttt aaacgcgttg ccataacttc gtataggata
     2701 ctttatacga agttatctgc atgctaggtt gccgggaacg gtgcattgga acgcgcattc
     2761 cccgtgttaa ttaacaggta agtgtcttcc tcctgtttcc ttcccctgct attctgctca
     2821 accttcctat cagaaactgc agtatctgta tttttgctag gggatcccgc caccatgggc
     2881 tctgaactga ttaaggagaa catgcacatg aagctgtaca tggagggaac tgtggataac
     2941 catcatttca agtgtaccag cgagggtgag ggtaagccct acgagggaac ccagactatg
     3001 agaattaaag ttgtggaggg aggtccgctg cctttcgcct ttgacatcct cgcgaccagc
     3061 ttcttgtacg gcagcaagac ttttatcaac cacactcaag gtatccctga cttttttaag
     3121 caaagttttc ctgagggatt cacctgggaa agagtaacca catacgaaga tgggggagtc
     3181 ctcacagcca ctcaagatac atcccttcaa gatggttgtt tgatctataa cgttaaaatc
     3241 cgcggagtga acttcaccag caacggaccc gttatgcaaa aaaaaactct cggatgggag
     3301 gccttcaccg agacgttgta tcctgcagat ggcggcttgg aagggcgcaa tgatatggcc
     3361 ctgaagctgg ttggcggtag tcatctgatc gctaatatca aaaccacata cagaagcaaa
     3421 aagccggcaa agaacctgaa gatgcctggt gtgtactacg ttgattaccg actggaaaga
     3481 attaaagagg ctaataacga gacatatgtg gagcagcacg aggtagccgt agccagatac
     3541 tgcgacctgc caagcaaact tgggcacaaa cttaactaga gcggcccctc tcgagcctct
     3601 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
     3661 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3721 ttgctttatt tgtaaccatt ataagctgca ataaacaaga gatctgcgaa gatacggcca
     3781 cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg ctgatgaatc
     3841 acatacttcc tccattttct tccactgatt gactgttata atttccctaa tttccaggtc
     3901 aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag gaccaatgaa
     3961 catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat aaataacatt
     4021 taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa
     4081 tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta aggagtaaaa
     4141 agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga tttttaattg
     4201 tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc aagtaaaatt
     4261 actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc cagcagatcc
     4321 actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg
     4381 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4441 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4501 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4561 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4621 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4681 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4741 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4801 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4861 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4921 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4981 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5041 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5101 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5161 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5221 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5281 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5341 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5401 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5461 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5521 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5581 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5641 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5701 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5761 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5821 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5881 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5941 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6001 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6061 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6121 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6181 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6241 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6301 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6361 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6421 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6481 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6541 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6601 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6661 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6721 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6781 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6841 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggg ctggctaaga
     6901 actcatcagc ctccccggtc catctaccca cgtaccaatg caccaattgg ccacaatgac
     6961 ggctactaca tggtgccatt ccttcctctt tataggaatg gagactacct cctgtccaac
     7021 aaggctcttg gatacgagta cgcctacctg ttggacccag gtcattgcac aacaccagaa
     7081 atgccctctg atctgcaaaa gacgtgaata tctgttcaga cacccatatc cactctgttc
     7141 cacacaggtc agaggtttgt ccaggagttc ttgacagagg tgtaaaaagt actcaaaaat
     7201 tttactcaag tgaaagtaca agtacttagg gaaaatttta ctcaattaaa agtaaaagta
     7261 tctggctaga atcttacttg agtaaaagta aaaaagtact ccattaaaat tgtacttgag
     7321 tattaaggaa gtaaaagtaa aagcaagaaa gaaaactaga gattcttgtt taagctttta
     7381 atctcaaaaa acattaaatg aaatgcatac aaggttttat cctgctttag aactgtttgt
     7441 atttaattat caaactataa gacagacaat ctaatgccag tacacgctac tcaaagttgt
     7501 aaaacctcag atttaacttc agtagaagct gattctcaaa attgttagtg tcaagcctag
     7561 ctcttttggg gctgaaaagc aatcctgcag tgctgaaaag cctctcacag gcagccgatg
     7621 cgggaagagg tgtattagtc ttgatagaga ggctgcaaat agcaggaaac gtgagcagag
     7681 actccctggt gtctgaaaca caggccagat gggccctcga g
//
