LOCUS       pKG2215_pGEEC525        3169 bp    DNA     circular SYN 02-MAR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3169)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..42
                     /label=oMJ001
     primer_bind     1..42
                     /label=oMJ005
     primer_bind     3..42
                     /label=oMJ009
     primer_bind     3..42
                     /label=oMJ013
     primer_bind     9..39
                     /label=oMJ020
     primer_bind     complement(21..46)
                     /label=oSEQ033 Pentr_rev
     primer_bind     complement(21..46)
                     /label=?oSEQ033 Pentr_rev
     misc_feature    23..46
                     /label=pENTR2B.Seq.REV
     primer_bind     complement(35..56)
                     /label=oTA069 mIR-backbone_junction_rev
     primer_bind     complement(35..56)
                     /label=oSEQ044 pENTR_rev_seq
     primer_bind     complement(108..128)
                     /label=oSEQ005 pENTR_rev
     CDS             122..931
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(147..168)
                     /label=oSEQ023 attl2_rev
     primer_bind     complement(155..178)
                     /label=Kan_rev_SEQ
     primer_bind     complement(155..178)
                     /label=OSEQ087 Kan_rev_SEQ
     primer_bind     complement(545..574)
                     /label=oTA099 GG_entry_upstream_fwd
     primer_bind     545..574
                     /label=oTA102 GG_entry_downstream_rev
     misc_feature    1005..1624
                     /label=pBR322_Origin
     rep_origin      1021..1609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1416..1431
                     /label=oTA073 Xho_addition_rev
     primer_bind     complement(1486..1501)
                     /label=oTA072 Xho_addition_for
     primer_bind     1510..1529
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     1604..1622
                     /label=ZF43_6x-C-fwd
     terminator      1774..1860
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1923..1942
                     /label=oSEQ011 pEntr_for
     terminator      1952..1979
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1981..2000
                     /label=pENTR_fwd_seq
     primer_bind     1981..2000
                     /label=oSEQ043 pENTR_fwd_seq
     primer_bind     complement(2002..2046)
                     /label=oMJ002
     primer_bind     complement(2002..2042)
                     /label=oMJ010
     primer_bind     complement(2002..2038)
                     /label=oMJ006
     primer_bind     complement(2002..2038)
                     /label=oMJ014
     primer_bind     complement(2010..2038)
                     /label=oMJ019
     primer_bind     2043..2067
                     /label=oEP054
     misc_feature    2047..2209
                     /label=ZF43_6x_C-promoter
     primer_bind     2047..2064
                     /label=ZF43_6x-C-fwd
     protein_bind    2065..2073
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2080..2088
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2095..2103
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2110..2118
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2125..2133
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     protein_bind    2140..2148
                     /label=ZF43 Binding Site
                     /bound_moiety="ZF43"
     primer_bind     complement(2189..2209)
                     /label=ZF43_6x_C-rev
     primer_bind     complement(2190..2213)
                     /label=oEP055
     regulatory      2214..2223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2217..2238
                     /label=GG_mRuby2_fwd
     primer_bind     2219..2238
                     /label=mRuby2_fwd
                     /note="Fwd primer for mRuby2"
     CDS             2220..2930
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     2220..2239
                     /label=oEP159
     primer_bind     complement(2246..2267)
                     /label=oEP076
     primer_bind     2268..2287
                     /label=oEP073
     primer_bind     2290..2309
                     /label=oSEQ062 mRuby2_fwd
     primer_bind     complement(2290..2309)
                     /label=oSEQ093 mRuby2_rev
     primer_bind     complement(2822..2841)
                     /label=oSEQ061 mRuby2_rev
     primer_bind     2856..2875
                     /label=OSEQ091 mRuby2_fwd_SEQ
     primer_bind     complement(2911..2931)
                     /label=oEP160
     primer_bind     complement(2915..2942)
                     /label=GG_mRuby2_rev
     primer_bind     complement(2915..2937)
                     /label=gemRuby2_rev
     primer_bind     complement(2915..2934)
                     /label=mRuby2_rev
                     /note="Rev primer for mRuby2"
     primer_bind     2933..2957
                     /label=gebGH_fwd
     primer_bind     2939..2960
                     /label=GG_bGH_fwd
     primer_bind     2943..2960
                     /label=GG_WPRE-bGH_fwd
     polyA_signal    2945..3169
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(join(3151..3169,1..4))
                     /label=gebGH_rev
     primer_bind     complement(3152..3168)
                     /label=oSEQ078 bGH_rev
     primer_bind     complement(3155..3169)
                     /label=GG_bGH_rev
ORIGIN
        1 cgctcgctga ctgcaggtgc ccgtgtctca aaatctctga tgttacattg cacaagataa
       61 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
      121 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
      181 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
      241 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
      301 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
      361 gaccatcaag cattttatcc gtactcctgg tgatgcatgg ttactcacca ctgcgatccc
      421 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
      481 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
      541 cagcgatcgc gtatttcgtc tagctcaggc gcaatcacga atgaataacg gtttggttga
      601 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
      661 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
      721 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
      781 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
      841 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
      901 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta
      961 ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1681 gttatcccct gattctgtgg ataaccgtat taccgctagc atggatctcg gggacgtcta
     1741 actactaagc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
     1801 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1861 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
     1921 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
     1981 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
     2041 cttactagat ctgaattacg atcggagtga ggataattag agtgaggacg atcggagtga
     2101 ggaaaattag agtgaggacg atcggagtga ggataattag agtgaggacg atccctctag
     2161 agggtatata atgggggcca ctagtctact accagagctc atcgctagca atggccacca
     2221 tggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg tatgaaggtg gtcatggaag
     2281 gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg agaaggcaat ccgtacatgg
     2341 gaactcaaac catgaggatc aaagtcatcg agggaggacc cctgccattt gcctttgaca
     2401 ttcttgccac gtcgttcatg tatggcagcc gtacttttat caagtacccg aaaggcattc
     2461 ctgatttctt taaacagtcc tttcctgagg gttttacttg ggaaagagtt acgagatacg
     2521 aagatggtgg agtcgtcacc gtcatgcagg acaccagcct tgaggatggc tgtctcgttt
     2581 accacgtcca agtcagaggg gtaaactttc cctccaatgg tcccgtgatg cagaagaaga
     2641 ccaagggttg ggagcctaat acagagatga tgtatccagc agatggtggt ctgaggggat
     2701 acactcatat ggcactgaaa gttgatggtg gtggccatct gtcttgctct ttcgtaacaa
     2761 cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc cggtatccat gccgttgatc
     2821 accgcctgga aaggttagag gaaagtgaca atgaaatgtt cgtagtacaa cgcgaacacg
     2881 cagttgccaa gttcgccggg cttggtggtg ggatggacga gctgtacaag taacaacgtt
     2941 gcacctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3001 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3061 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3121 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatgg
//
