LOCUS       Exported                7544 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 7544)
  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..7544
                     /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     complement(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(1748..1764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1780..1914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1915..2634)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     promoter        complement(2641..3695)
                     /label=exorh
                     /label=PIPE
     misc_recomb     complement(3697..4031)
                     /label=Tol2 exon1 bp1-335 of 461
     LTR             4195..4213
                     /label=Tol2 3'TIR
     promoter        complement(4300..4318)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4334..4355)
                     /label=M13R
     primer_bind     complement(4339..4355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4741..5329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5500..6360)
                     /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             6998..7014
                     /label=Tol2 3' TIR
     misc_structure  7187..7533
                     /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 tccatgtcat gtgtaatccc ggcggcggtc accctctcct tcaggaccat
     1981 gtgatcgcgc ttctcgttgg ggtctttgct cagtttggac tgatggctca ggtagtggtt
     2041 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     2101 ctgcacgccg ccgtcctcaa cgttgtggcg ggtcttgaag ttagccttga tgccgttctt
     2161 ctgcttgtcg gccgtgatat agaccttgtg gctgttgaag ttgtactcca gcttgtgccc
     2221 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc acgatgcggt tcaccagggt
     2281 gtcgccctcg aacttcacct cggcgcgggt cttgtaggta ccgtcgtcct tgaaagagat
     2341 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     2401 gtggtcgggg tagcgggcga agcaggccac gccgtagcct aaggtggtca cgagggtggg
     2461 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgttggt
     2521 ggcatcgccc tcgccctcgc cgcggacgct gaacttgtgg ccgtttacgt cgccgtccag
     2581 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     2641 gatggagaag tggacgatcg gcacacctcc gcagatgaag cccggagagc cggcgctgcg
     2701 ctcttatgtg aaccccatga gtccagatta ggcagattac aggagtgagt ggtcagattg
     2761 gggtcagaca gggggataat ccagagatgg agacacagca gcgctcgctc tcaccacaca
     2821 cactcacgac acacacacac acactactgc tgctcatcag cgcgtgcagg ctgcgtctcc
     2881 atccaaacac tcctgcacag agctgaaatc tctcatccac agacagacag gtgtcctgca
     2941 gaacacaacc ctcctgctgg acaacacaca cacacacaca cacacacaca cacacacaca
     3001 cacacacaca cacagcaagt cctgtgtgag cctcataaga tgaagaccaa acacaaggga
     3061 cacttcacgg tctcacacac acacacacac acacacacac acacacacac acacacacac
     3121 acacacacac acacacacag attatcagac tgaaatatta aatctttatt aaaaacacca
     3181 acatgaggga tgtgtggatg cgcaggagtg tgttgagctg tgagtgtgtg tgtgtgtgtg
     3241 tgtgtgtgtg tccagcatcg ctgcaatctg ctttgtgttg atctgtgtaa tcctatcaga
     3301 gctgtgctga gctggcattt actgacggct ttacagcggc tcagggtgga tcacaccgcg
     3361 gtaaacatga gtgtgtgtta gcaggactga tgatcacacc gcggtaaaca tgagtgtgtg
     3421 ttagcaggac tcatcgatgg gtctgcggca gaacgggcag cagctgtgct gaagaaccaa
     3481 cagctcatag tcctcactgt ggaacatctg agaaacacac acacacacac acacacacaa
     3541 ccatcagaca cacaacaaag catatagagt caaacagaca aataacagag gagacacaca
     3601 cacacacaca cacagatgag cgcacctgga agcaggatgg gcacatggtg atgctgacgt
     3661 cgggcaggag tgagcggtag tactgccagc tgagctgcga agatacggcc acgggtgctc
     3721 ttgatcctgt ggctgatttt ggactgtgct gctcgcagct gctgatgaat cacatacttc
     3781 ctccattttc ttccactgat tgactgttat aatttcccta atttccaggt caaggtgctg
     3841 tgcattgtgg taatagatgt gacatgacgt cacttccaaa ggaccaatga acatgtctga
     3901 ccaatttcat ataatgtgaa aacgattttc ataggcagaa taaataacat ttaaattaaa
     3961 ctgggcatca gcgcaattca attggtttgg taatagcaag ggaaaataga atgaagtgat
     4021 ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt
     4081 ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt
     4141 aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta
     4201 ctttacacct ctggttcttg accccctacc ttcagcaagc ccagcagatc cactagttct
     4261 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc
     4321 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5161 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6481 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6541 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6601 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6661 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc acgctggcta
     6721 agaactcatc agcctccccg gtccatctac ccacgtacca atgcaccaat tggccacaat
     6781 gacggctact acatggtgcc attccttcct ctttatagga atggagacta cctcctgtcc
     6841 aacaaggctc ttggatacga gtacgcctac ctgttggacc caggtcattg cacaacacca
     6901 gaaatgccct ctgatctgca aaagacgtga atatctgttc agacacccat atccactctg
     6961 ttccacacag gtcagaggtt tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa
     7021 aattttactc aagtgaaagt acaagtactt agggaaaatt ttactcaatt aaaagtaaaa
     7081 gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt actccattaa aattgtactt
     7141 gagtattaag gaagtaaaag taaaagcaag aaagaaaact agagattctt gtttaagctt
     7201 ttaatctcaa aaaacattaa atgaaatgca tacaaggttt tatcctgctt tagaactgtt
     7261 tgtatttaat tatcaaacta taagacagac aatctaatgc cagtacacgc tactcaaagt
     7321 tgtaaaacct cagatttaac ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc
     7381 tagctctttt ggggctgaaa agcaatcctg cagtgctgaa aagcctctca caggcagccg
     7441 atgcgggaag aggtgtatta gtcttgatag agaggctgca aatagcagga aacgtgagca
     7501 gagactccct ggtgtctgaa acacaggcca gatgggccct cgag
//
