LOCUS       Exported                4234 bp ds-DNA     circular SYN 21-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4234)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 10, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4234
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      269..296
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      388..474
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     538..554
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..669
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     670..687
                     /label=M13-fwd(1)
     primer_bind     671..687
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        697..715
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    724..745
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     741..745
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     745..788
                     /label=oJDW 1950 LiCre FWD + Kozak + SalI
     primer_bind     751..788
                     /label=oJDW 1939 LiCre FWD + Kozak + AgeI + attB1
     regulatory      751..760
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             757..2228
                     /codon_start=1
                     /label=LiCre
                     /translation="MGPKKKRKVDPKKKRKVMRGSHHHHHHGSLATTLERIEKNFVITD
                     PRLPDNPIIFASDSFLQLTEYSREEILGRNCRFLQGPETDRATVRKIRDAIDNQTEVTV
                     QLINYTKSGKKFWNVFHLQPMRDYKGDVQYFIGVQLDGTERLHGAAEREAVCLIKKTAF
                     QIDSDEVRKNLMDMFRDRQAFSEHTWKMLLSVCRSWAAWCKLNNRKWFPAEPEDVRDYL
                     LYLQARGLAVKTIQQHLGQLNMLHRRSGLPRPSDSNAVSLVMRRIRKENVDAGERAKQA
                     LAFERTDFDQVRSLMENSDRCQDIRNLAFLGIAYNTLLRIAEIARIRVKDISRTDGGRM
                     LIHIGRTKTLVSTAGVEKALSLGVTKLVERWISVSGVADDPNNYLFCRVRKNGVAAPSA
                     TSQLSTRALEGIFEATHRLIYGAKDDSGQRYLAWSGHSARVGAARDMARAGVSIPEIMQ
                     AGGWTNVNIVMNYIRNLDSETGAMVRLLAAGD"
     CDS             763..783
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             787..807
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             820..837
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2229..2299
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(2259..2275)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2312..2330)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(2359..2458)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2476..2494)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2499..2515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2628..3437
                     /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      3584..4172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
       61 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      121 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      181 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      241 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      301 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      361 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      421 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      481 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      541 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      601 cctgttcgtt gcaacacatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      661 aagcaggctt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
      721 ttgggtaccg ggccccccct cgaggtcgac gccaccatgg gtccaaaaaa gaagagaaag
      781 gtagatccaa aaaagaagag aaaggtaatg aggggcagcc accaccacca ccaccacggc
      841 agcctggcca ccaccctgga gaggatcgag aagaacttcg tgatcaccga ccccaggctg
      901 cccgacaacc ccatcatctt cgccagcgac agcttcctgc agctgaccga gtacagcagg
      961 gaggagatcc tgggcaggaa ctgcaggttc ctgcagggcc ccgagaccga cagggccacc
     1021 gtgaggaaga tcagggacgc catcgacaac cagaccgagg tgaccgtgca gctgatcaac
     1081 tacaccaaga gcggcaagaa gttctggaac gtgttccacc tgcagcccat gagggactac
     1141 aagggcgacg tgcagtactt catcggcgtg cagctggacg gcaccgagag gctgcacggc
     1201 gccgccgaga gggaggccgt gtgcctgatc aagaagaccg ccttccagat cgacagcgac
     1261 gaggtgagga agaacctgat ggacatgttc agggacaggc aggccttcag cgagcacacc
     1321 tggaagatgc tgctgagcgt gtgcaggagc tgggccgcct ggtgcaagct gaacaacagg
     1381 aagtggttcc ccgccgagcc cgaggacgtg agggactacc tgctgtacct gcaggccagg
     1441 ggcctggccg tgaagaccat ccagcagcac ctgggccagc tgaacatgct gcacaggagg
     1501 agcggcctgc ccaggcccag cgacagcaac gccgtgagcc tggtgatgag gaggatcagg
     1561 aaggagaacg tggacgccgg cgagagggcc aagcaggccc tggccttcga gaggaccgac
     1621 ttcgaccagg tgaggagcct gatggagaac agcgacaggt gccaggacat caggaacctg
     1681 gccttcctgg gcatcgccta caacaccctg ctgaggatcg ccgagatcgc caggatcagg
     1741 gtgaaggaca tcagcaggac cgacggcggc aggatgctga tccacatcgg caggaccaag
     1801 accctggtga gcaccgccgg cgtggagaag gccctgagcc tgggcgtgac caagctggtg
     1861 gagaggtgga tcagcgtgag cggcgtggcc gacgacccca acaactacct gttctgcagg
     1921 gtgaggaaga acggcgtggc cgcccccagc gccaccagcc agctgagcac cagggccctg
     1981 gagggcatct tcgaggccac ccacaggctg atctacggcg ccaaggacga cagcggccag
     2041 aggtacctgg cctggagcgg ccacagcgcc agggtgggcg ccgccaggga catggccagg
     2101 gccggcgtga gcatccccga gatcatgcag gccggcggct ggaccaacgt gaacatcgtg
     2161 atgaactaca tcaggaacct ggacagcgag accggcgcca tggtgaggct gctggccgcc
     2221 ggcgactaag cttgatatcg aattcctgca gcccggggga tccactagtt ctagagcggc
     2281 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     2341 cgtaatcatg gtcatagcac ccagctttct tgtacaaagt tggcattata agaaagcatt
     2401 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2461 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2521 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2581 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2641 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2701 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     2761 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2821 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2881 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2941 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3001 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3061 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3121 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3181 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3241 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3301 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3361 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3421 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3481 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3541 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3601 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3661 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3721 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3781 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3841 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3901 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3961 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4021 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4081 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4141 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4201 tcctggcctt ttgctggcct tttgctcaca tgtt
//
