LOCUS       Exported                3098 bp ds-DNA     circular SYN 23-FEB-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3098)
  AUTHORS   Baylor-NRI
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3098
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     terminator      218..245
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      337..423
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     487..503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    543..638
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        640..658
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    667..682
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        685..1034
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    693..711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    729..747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    765..783
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    801..819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    837..855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    873..891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    909..927
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    929..940
                     /label=pTight Hybrid Sequence
     promoter        join(937,942..1004)
                     /label=Modified Minimal CMV promoter (pTRE3G Clontech)
     misc_feature    1042..1112
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(1072..1088)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1125..1143)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1143..1267
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1340..1358)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1363..1379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1492..2301
                     /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      2448..3036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
       61 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      121 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
      181 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
      241 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
      301 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
      361 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
      421 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
      481 gacgttgtaa aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct
      541 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca
      601 atgctttttt ataatgccaa ctttgtatag aaaagttggt aatacgactc actatagggc
      661 gaattgggta ccgggccccc cctcgagttt actccctatc agtgatagag aacgtatgaa
      721 gagtttactc cctatcagtg atagagaacg tatgcagact ttactcccta tcagtgatag
      781 agaacgtata aggagtttac tccctatcag tgatagagaa cgtatgacca gtttactccc
      841 tatcagtgat agagaacgta tctacagttt actccctatc agtgatagag aacgtatatc
      901 cagtttactc cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggcgc
      961 ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt ccacaacact
     1021 tttgtcttat acttacgcgt aagcttgata tcgaattcct gcagcccggg ggatccacta
     1081 gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     1141 attcaagttt gtacaaaaaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
     1201 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatgcagt
     1261 cactatgaac caactactta gatggtatta gtgacctgta gaattcgagc tctagagctg
     1321 cagggcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     1381 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1441 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1501 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1561 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1621 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1681 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1741 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1801 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1861 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1921 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1981 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     2041 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2101 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2161 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2221 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2281 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2341 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     2401 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2461 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2521 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2581 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2641 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2701 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2761 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2821 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2881 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2941 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3001 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3061 cggttcctgg ccttttgctg gccttttgct cacatgtt
//
