LOCUS       Exported                3974 bp ds-DNA     circular SYN 14-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3974)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3974
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    689..697
                     /label=Modified Kozak
     CDS             698..1489
                     /codon_start=1
                     /label=TAEL2.0 (EL222_c.o.)
                     /translation="MLPADALDDFDLDMLPADALDDYDLDILPGNEFGADDTRVEVQPP
                     AQWVLDLIEASPIASVVSDPRLADNPLIAINQAFTDLTGYSEEECVGRNCRFLAGSGTE
                     PWLTDKIRQGVREHKPVLVEILNYKKDGTPFRNAVLVAPIYDDDDELLYFLGSQVEVDD
                     DQPNMGMARRERAAEMLKTLSPRQLEVTTLVASGLRNKEVAARLGLSEKTVKMHRGLVM
                     EKLNLKTSADLVRIAVEAGIEFSASKRPAATKKAGQAKKKK"
     CDS             698..772
                     /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(704..726)
                     /label=oJDW 1984 (TAEL2.0 REV SEQ)
     CDS             1439..1486
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     1496..1537
                     /label=oJDW XXXX (p5E FWD)
     misc_feature    1507..1698
                     /locus_tag="SV40 polyA"
                     /label=SV40 polyA
     misc_feature    1581..1600
                     /label=SV40pA REVSeq
     misc_feature    1711..1802
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1751..1770
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     misc_feature    1816..2062
                     /label=NOS terminator
                     /note="nopaline synthase terminator from Agrobacterium
                     tumefaciens (NOS terminator)"
     protein_bind    complement(2099..2198)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2239..2255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2368..3177
                     /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"
     rep_origin      3324..3912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg atccactagt ttaattaagc cgccaccatg ttgcctgcag atgcacttga
      721 tgactttgac ctggacatgc tcccagctga tgctctggat gactatgacc tcgacatcct
      781 gcctgggaat gagtttggag cggacgacac tcgtgttgaa gtgcagccac ctgcccagtg
      841 ggtgctcgat ctcatagaag caagtccgat tgcatctgtg gtgtccgacc ctcgcttagc
      901 agacaatcca cttattgcca tcaaccaggc cttcacagat ttaaccggct acagtgagga
      961 agagtgtgtg ggtaggaact gccgctttct ggccgggagc ggaacagagc cctggctgac
     1021 tgacaagatc aggcaaggag tcagagagca taaaccagta ttggttgaga tcctcaacta
     1081 caagaaagat ggaacgccat tcagaaatgc tgttctagta gctcccatct atgatgatga
     1141 tgatgagctg ctgtacttcc tgggctctca ggtggaggtg gacgatgacc agcccaacat
     1201 gggcatggcc agacgagaac gagccgcaga gatgctgaag acactatcgc cgcggcagct
     1261 ggaggtcacc acccttgtgg ctagcgggtt gcgcaacaaa gaagttgcag cgcggttggg
     1321 tctttcagaa aagactgtca agatgcacag aggtctcgtc atggagaaat tgaatctgaa
     1381 aacatctgct gatctggtca ggatagctgt ggaagccggc attgagttct ccgcgtcaaa
     1441 gagacctgct gccactaaaa aagctggaca agccaagaag aaaaaatgat aaccatggaa
     1501 ttctagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1561 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1621 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1681 aggtgtggga ggttttttat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     1741 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     1801 ctcctcgaga gatctttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     1861 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     1921 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     1981 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     2041 gaattgatcc cccctcgaca gcggtaccac gcgtacgcgt gaattcgata tcaagcttac
     2101 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2161 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     2221 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2281 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2341 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2401 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2461 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2521 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2581 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2641 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2701 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2761 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2821 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2881 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2941 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     3001 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     3061 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     3121 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     3181 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     3241 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3301 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3361 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3421 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3481 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3541 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3601 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3661 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3721 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3781 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3841 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3901 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3961 tttgctcaca tgtt
//
