LOCUS       pKG3779_pGEEC586        3561 bp    DNA     circular SYN 23-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3561)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3561
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     10..29
                     /label=pENTR_fwd_seq
     primer_bind     10..29
                     /label=oSEQ043 pENTR_fwd_seq
     primer_bind     complement(31..75)
                     /label=oMJ002
     primer_bind     complement(31..71)
                     /label=oMJ010
     primer_bind     complement(31..67)
                     /label=oMJ006
     primer_bind     complement(31..67)
                     /label=oMJ014
     primer_bind     complement(39..67)
                     /label=oMJ019
     primer_bind     72..98
                     /label=pPV1-CMV_fwd
     primer_bind     72..93
                     /label=oEP143
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(247..266)
                     /label=oSEQ065 CMV_rev
     promoter        380..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     537..557
                     /label=oSEQ030 CMV_fwd
     primer_bind     complement(552..571)
                     /label=oSEQ042 SFCp_rev
     primer_bind     complement(565..603)
                     /label=pPV1-CMV_rev
     primer_bind     complement(569..588)
                     /label=oEP138
     primer_bind     complement(569..588)
                     /label=oEP144
     primer_bind     584..601
                     /label=pPV1-EF1a-bb_fwd
     primer_bind     complement(585..630)
                     /label=oEP202
     primer_bind     585..604
                     /label=oEP206 CMV-rev
     primer_bind     605..650
                     /label=oEP201
     misc_feature    605..630
                     /label=EFS 5' UTR
     primer_bind     608..627
                     /label=oEP209 EF1a-rev
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     638..659
                     /label=GG_mRuby2_fwd
     primer_bind     640..659
                     /label=mRuby2_fwd
                     /note="Fwd primer for mRuby2"
     CDS             641..1351
                     /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     641..660
                     /label=oEP159
     primer_bind     complement(667..688)
                     /label=oEP076
     primer_bind     689..708
                     /label=oEP073
     primer_bind     711..730
                     /label=oSEQ062 mRuby2_fwd
     primer_bind     complement(711..730)
                     /label=oSEQ093 mRuby2_rev
     primer_bind     complement(1243..1262)
                     /label=oSEQ061 mRuby2_rev
     primer_bind     1277..1296
                     /label=OSEQ091 mRuby2_fwd_SEQ
     primer_bind     complement(1332..1352)
                     /label=oEP160
     primer_bind     complement(1336..1363)
                     /label=GG_mRuby2_rev
     primer_bind     complement(1336..1358)
                     /label=gemRuby2_rev
     primer_bind     complement(1336..1355)
                     /label=mRuby2_rev
                     /note="Rev primer for mRuby2"
     primer_bind     1354..1378
                     /label=gebGH_fwd
     primer_bind     1360..1381
                     /label=GG_bGH_fwd
     primer_bind     1364..1381
                     /label=GG_WPRE-bGH_fwd
     polyA_signal    1366..1590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1450..1475)
                     /label=oEP181 IVT-bGH-rev
     misc_feature    1473..1475
                     /label=bGH TES
     primer_bind     complement(1572..1594)
                     /label=gebGH_rev
     primer_bind     complement(1573..1589)
                     /label=oSEQ078 bGH_rev
     primer_bind     complement(1576..1590)
                     /label=GG_bGH_rev
     primer_bind     1591..1632
                     /label=oMJ001
     primer_bind     1591..1632
                     /label=oMJ005
     primer_bind     1593..1632
                     /label=oMJ009
     primer_bind     1593..1632
                     /label=oMJ013
     primer_bind     1599..1629
                     /label=oMJ020
     primer_bind     complement(1611..1636)
                     /label=oSEQ033 Pentr_rev
     misc_feature    1613..1636
                     /label=pENTR2B.Seq.REV
     primer_bind     complement(1625..1646)
                     /label=oTA069 mIR-backbone_junction_rev
     primer_bind     complement(1625..1646)
                     /label=oSEQ044 pENTR_rev_seq
     primer_bind     complement(1698..1718)
                     /label=oSEQ005 pENTR_rev
                     /note="25nm	STD"
     CDS             1712..2521
                     /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(1737..1758)
                     /label=oSEQ023 attl2_rev
     primer_bind     complement(1745..1768)
                     /label=Kan_rev_SEQ
     primer_bind     complement(2135..2164)
                     /label=oTA099 GG_entry_upstream_fwd
     primer_bind     2135..2164
                     /label=oTA102 GG_entry_downstream_rev
     misc_feature    2595..3214
                     /label=pBR322_Origin
     rep_origin      2611..3199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2643..2662
                     /label=oEP033
     primer_bind     complement(2643..2662)
                     /label=oEP035
     primer_bind     2963..2983
                     /label=oEP024
     primer_bind     complement(2963..2983)
                     /label=oEP025
     primer_bind     3006..3021
                     /label=oTA073 Xho_addition_rev
     primer_bind     complement(3076..3091)
                     /label=oTA072 Xho_addition_for
     primer_bind     3100..3119
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     complement(3216..3235)
                     /label=oEP013
     primer_bind     3216..3235
                     /label=oEP014
     primer_bind     3351..3364
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      3364..3450
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3513..3532
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      join(3542..3561,1..8)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggcccacc
       61 tgcagtctac ttactcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctgagctcg tttagtgaac
      601 cgtccaacgg gtttgccgcc agaacacagt aatggccacc atggtgtcta agggcgaaga
      661 gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca
      721 attcaaatgc acaggtgaag gagaaggcaa tccgtacatg ggaactcaaa ccatgaggat
      781 caaagtcatc gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat
      841 gtatggcagc cgtactttta tcaagtaccc gaaaggcatt cctgatttct ttaaacagtc
      901 ctttcctgag ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac
      961 cgtcatgcag gacaccagcc ttgaggatgg ctgtctcgtt taccacgtcc aagtcagagg
     1021 ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa
     1081 tacagagatg atgtatccag cagatggtgg tctgagggga tacactcata tggcactgaa
     1141 agttgatggt ggtggccatc tgtcttgctc tttcgtaaca acttacaggt caaaaaagac
     1201 cgtcgggaac atcaagatgc ccggtatcca tgccgttgat caccgcctgg aaaggttaga
     1261 ggaaagtgac aatgaaatgt tcgtagtaca acgcgaacac gcagttgcca agttcgccgg
     1321 gcttggtggt gggatggacg agctgtacaa gtaacaacgt tgcacctgtg ccttctagtt
     1381 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1441 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1501 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     1561 ggcatgctgg ggatgcggtg ggctctatgg cgctcgctga ctgcaggtgc ccgtgtctca
     1621 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1681 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1741 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1801 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     1861 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     1921 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctgg
     1981 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2041 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2101 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tagctcaggc
     2161 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2221 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2281 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2341 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2401 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2461 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2521 atcagaattg gttaattggt tgtaacatta ttcagattgg gccccgttcc actgagcgtc
     2581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3301 taccgctagc atggatctcg gggacgtcta actactaagc gagagtaggg aactgccagg
     3361 catcaaataa aacgaaaggc tcagtcggaa gactgggcct ttcgttttat ctgttgtttg
     3421 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgtgaag
     3481 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaactaag
     3541 cagaaggcca tcctgacgga t
//
