LOCUS       Exported                4148 bp ds-DNA     circular SYN 27-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4148)
  AUTHORS   Wythe Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4148
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1919..1926
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    189..288
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    303..311
                     /label=Modified Kozak
     CDS             312..1103
                     /codon_start=1
                     /label=TAEL2.0 (EL222_c.o.)
                     /translation="MLPADALDDFDLDMLPADALDDYDLDILPGNEFGADDTRVEVQPP
                     AQWVLDLIEASPIASVVSDPRLADNPLIAINQAFTDLTGYSEEECVGRNCRFLAGSGTE
                     PWLTDKIRQGVREHKPVLVEILNYKKDGTPFRNAVLVAPIYDDDDELLYFLGSQVEVDD
                     DQPNMGMARRERAAEMLKTLSPRQLEVTTLVASGLRNKEVAARLGLSEKTVKMHRGLVM
                     EKLNLKTSADLVRIAVEAGIEFSASKRPAATKKAGQAKKKK"
     CDS             312..386
                     /codon_start=1
                     /label=T4A
                     /note="transactivating domain of KalTA4, which has been 
                     reported to be better tolerated in zebrafish embryos (Baron
                     et al., 1997; Distel et al., 2009) "
                     /translation="MLPADALDDFDLDMLPADALDDYDL"
     primer_bind     complement(318..340)
                     /label=oJDW 1984 (TAEL2.0 REV SEQ)
     CDS             1053..1100
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     1110..1151
                     /label=oJDW XXXX (p5E FWD)
     misc_feature    1121..1312
                     /locus_tag="SV40 polyA"
                     /label=SV40 polyA
     misc_feature    1195..1214
                     /label=SV40pA REVSeq
     misc_feature    1325..1416
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1365..1384
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     misc_feature    1430..1676
                     /label=NOS terminator
                     /note="nopaline synthase terminator from Agrobacterium
                     tumefaciens (NOS terminator)"
     misc_feature    1696..1815
                     /label=C120 regulatory
     misc_feature    1696..1715
                     /label=20bp version of Clone-1
     misc_feature    1721..1740
                     /label=20bp version of Clone-1
                     /label=20bp version of Clone-1(1)
     misc_feature    1746..1765
                     /label=20bp version of Clone-1
                     /label=20bp version of Clone-1(2)
     misc_feature    1771..1790
                     /label=20bp version of Clone-1
                     /label=20bp version of Clone-1(3)
     misc_feature    1796..1815
                     /label=20bp version of Clone-1
                     /label=20bp version of Clone-1(4)
     promoter        1823..1915
                     /label=mmu-cFos min pro
                     /label=cFos
                     /note="murine c-Fos minimal promoter"
     primer_bind     1857..1879
                     /label=oJDW 1408 (c-fos min pro FWD)
     primer_bind     complement(1897..1915)
                     /label=oJDW 1983 (c-fos min pro REV)
     primer_bind     complement(1910..1938)
                     /label=oJDW XXX (p5E REV)
     protein_bind    complement(1940..2039)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2058..2074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2082..2098
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2106..2136)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2151..2172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2460..3048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3226..4035)
                     /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"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacacattg
      241 atgagcaatg cttttttata atgccaactt tgtacaaaaa agcaggctgg atccttaatt
      301 aagccgccac catgttgcct gcagatgcac ttgatgactt tgacctggac atgctcccag
      361 ctgatgctct ggatgactat gacctcgaca tcctgcctgg gaatgagttt ggagcggacg
      421 acactcgtgt tgaagtgcag ccacctgccc agtgggtgct cgatctcata gaagcaagtc
      481 cgattgcatc tgtggtgtcc gaccctcgct tagcagacaa tccacttatt gccatcaacc
      541 aggccttcac agatttaacc ggctacagtg aggaagagtg tgtgggtagg aactgccgct
      601 ttctggccgg gagcggaaca gagccctggc tgactgacaa gatcaggcaa ggagtcagag
      661 agcataaacc agtattggtt gagatcctca actacaagaa agatggaacg ccattcagaa
      721 atgctgttct agtagctccc atctatgatg atgatgatga gctgctgtac ttcctgggct
      781 ctcaggtgga ggtggacgat gaccagccca acatgggcat ggccagacga gaacgagccg
      841 cagagatgct gaagacacta tcgccgcggc agctggaggt caccaccctt gtggctagcg
      901 ggttgcgcaa caaagaagtt gcagcgcggt tgggtctttc agaaaagact gtcaagatgc
      961 acagaggtct cgtcatggag aaattgaatc tgaaaacatc tgctgatctg gtcaggatag
     1021 ctgtggaagc cggcattgag ttctccgcgt caaagagacc tgctgccact aaaaaagctg
     1081 gacaagccaa gaagaaaaaa tgataaccat ggaattctag cagacatgat aagatacatt
     1141 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1201 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1261 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttatcgatag
     1321 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     1381 tccccagtgc aagtgcaggt gccagaacat ttctctcctc gagagatctt taagattgaa
     1441 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
     1501 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     1561 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     1621 atcgcgcgcg gtgtcatcta tgttactaga tcgggaattg atcccccctc gacagcggta
     1681 ccacgcgttg agctctaggt agcctttagt ccatgcgtta taggtagcct ttagtccatg
     1741 cgttataggt agcctttagt ccatgcgtta taggtagcct ttagtccatg cgttataggt
     1801 agcctttagt ccatgcgtcg acccagtgac gtaggaagtc catccattca cagcgcttct
     1861 ataaaggcgc cagctgaggc gcctactact ccaaccgcga ctgcagcgag caactaacgc
     1921 ggccgcacta gttctagact cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1981 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttga
     2041 agcttggtgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2881 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3181 tcaagcccaa tctgaataat gttacaacca attaaccaat tctgattaga aaaactcatc
     3241 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     3301 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     3361 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     3421 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     3481 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     3541 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     3601 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3661 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3721 gaatgctgtt tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3781 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3841 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3901 gggcttccca tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3961 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg
     4021 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     4081 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca
     4141 gagctgca
//
