LOCUS       Exported                7535 bp ds-DNA     circular SYN 13-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7535)
  AUTHORS   Stowers Institute
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 16, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7535
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..22
                     /label=M13R
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     37..160
                     /gene="mutant version of attR"
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="/bound_moiety=LR Clonase(TM)"
     misc_feature    complement(201..506)
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /label=nonstandard type:  Resistance
                     /note="Plasmids containing the ccdB gene cannot be
                     propagated in standard E. coli strains."
                     /note="/codon_start=1"
     CDS             complement(848..1507)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_recomb     1617..1740
                     /gene="mutant version of attR"
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="/bound_moiety=LR Clonase(TM)"
     primer_bind     complement(1749..1764)
                     /label=M13R (1)
     polyA_signal    1780..1914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1915..2625)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent
                     protein (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGHHSTGGMDELYK"
     promoter        complement(2632..3686)
                     /label=exorh
                     /label=PIPE
     misc_structure  complement(3688..4022)
                     /label=Tol2 exon1 bp1-335 of 461
     LTR             4186..4204
                     /label=Tol2 3'TIR
     promoter        complement(4291..4309)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4325..4346)
                     /label=M13R
     primer_bind     complement(4330..4346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4732..5320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5491..6351)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     LTR             6989..7005
                     /label=Tol2 3' TIR
     misc_structure  7178..7524
                     /label=Tol2 exon4 136-482
                     /label=tol2 gene exon 4
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 caaaggagcc
      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 agttttactt
     1921 gtacagctcg tccatgccgc cggtggagtg gtggccctcg gcgcgctcgt actgttccac
     1981 gatggtgtag tcctcgttgt gggaggtgat gtccagcttg atgttgacgt tgtaggcgcc
     2041 gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcggcgt cgtagtggcc
     2101 gccgtccttc agcttcagcc tctgcttgat ctcgcccttc agggcgccgt cctcggggta
     2161 catccgctcg gaggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcgga
     2221 ggggaagttg gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct gcagggagga
     2281 gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa
     2341 gccctcgggg aaggacagct tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc
     2401 cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca gggggccgcc
     2461 cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc
     2521 ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga agcgcatgaa
     2581 ctccttgatg atggccatgt tgtcatcctc gcccttgctc accatggtgg cgatggagaa
     2641 gtggacgatc ggcacacctc cgcagatgaa gcccggagag ccggcgctgc gctcttatgt
     2701 gaaccccatg agtccagatt aggcagatta caggagtgag tggtcagatt ggggtcagac
     2761 agggggataa tccagagatg gagacacagc agcgctcgct ctcaccacac acactcacga
     2821 cacacacaca cacactactg ctgctcatca gcgcgtgcag gctgcgtctc catccaaaca
     2881 ctcctgcaca gagctgaaat ctctcatcca cagacagaca ggtgtcctgc agaacacaac
     2941 cctcctgctg gacaacacac acacacacac acacacacac acacacacac acacacacac
     3001 acacagcaag tcctgtgtga gcctcataag atgaagacca aacacaaggg acacttcacg
     3061 gtctcacaca cacacacaca cacacacaca cacacacaca cacacacaca cacacacaca
     3121 cacacacaca gattatcaga ctgaaatatt aaatctttat taaaaacacc aacatgaggg
     3181 atgtgtggat gcgcaggagt gtgttgagct gtgagtgtgt gtgtgtgtgt gtgtgtgtgt
     3241 gtccagcatc gctgcaatct gctttgtgtt gatctgtgta atcctatcag agctgtgctg
     3301 agctggcatt tactgacggc tttacagcgg ctcagggtgg atcacaccgc ggtaaacatg
     3361 agtgtgtgtt agcaggactg atgatcacac cgcggtaaac atgagtgtgt gttagcagga
     3421 ctcatcgatg ggtctgcggc agaacgggca gcagctgtgc tgaagaacca acagctcata
     3481 gtcctcactg tggaacatct gagaaacaca cacacacaca cacacacaca accatcagac
     3541 acacaacaaa gcatatagag tcaaacagac aaataacaga ggagacacac acacacacac
     3601 acacagatga gcgcacctgg aagcaggatg ggcacatggt gatgctgacg tcgggcagga
     3661 gtgagcggta gtactgccag ctgagctgcg aagatacggc cacgggtgct cttgatcctg
     3721 tggctgattt tggactgtgc tgctcgcagc tgctgatgaa tcacatactt cctccatttt
     3781 cttccactga ttgactgtta taatttccct aatttccagg tcaaggtgct gtgcattgtg
     3841 gtaatagatg tgacatgacg tcacttccaa aggaccaatg aacatgtctg accaatttca
     3901 tataatgtga aaacgatttt cataggcaga ataaataaca tttaaattaa actgggcatc
     3961 agcgcaattc aattggtttg gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa
     4021 ataagtactt tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa
     4081 aaaatgtaat taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa
     4141 atccccaaaa ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc
     4201 tctggttctt gaccccctac cttcagcaag cccagcagat ccactagttc tagagcggcc
     4261 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     4321 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4381 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4441 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4561 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4621 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5161 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6481 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     6541 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     6601 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     6661 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cacgctggct aagaactcat
     6721 cagcctcccc ggtccatcta cccacgtacc aatgcaccaa ttggccacaa tgacggctac
     6781 tacatggtgc cattccttcc tctttatagg aatggagact acctcctgtc caacaaggct
     6841 cttggatacg agtacgccta cctgttggac ccaggtcatt gcacaacacc agaaatgccc
     6901 tctgatctgc aaaagacgtg aatatctgtt cagacaccca tatccactct gttccacaca
     6961 ggtcagaggt ttgtccagga gttcttgaca gaggtgtaaa aagtactcaa aaattttact
     7021 caagtgaaag tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc
     7081 tagaatctta cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattaa
     7141 ggaagtaaaa gtaaaagcaa gaaagaaaac tagagattct tgtttaagct tttaatctca
     7201 aaaaacatta aatgaaatgc atacaaggtt ttatcctgct ttagaactgt ttgtatttaa
     7261 ttatcaaact ataagacaga caatctaatg ccagtacacg ctactcaaag ttgtaaaacc
     7321 tcagatttaa cttcagtaga agctgattct caaaattgtt agtgtcaagc ctagctcttt
     7381 tggggctgaa aagcaatcct gcagtgctga aaagcctctc acaggcagcc gatgcgggaa
     7441 gaggtgtatt agtcttgata gagaggctgc aaatagcagg aaacgtgagc agagactccc
     7501 tggtgtctga aacacaggcc agatgggccc tcgag
//
