LOCUS       pKG3780_pGEEC587        3598 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 3598)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3598
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     1..20
                     /label=pENTR_fwd_seq
     primer_bind     1..20
                     /label=oSEQ043 pENTR_fwd_seq
     primer_bind     complement(22..66)
                     /label=oMJ002
     primer_bind     complement(22..62)
                     /label=oMJ010
     primer_bind     complement(22..58)
                     /label=oMJ006
     primer_bind     complement(22..58)
                     /label=oMJ014
     primer_bind     complement(30..58)
                     /label=oMJ019
     primer_bind     63..89
                     /label=pPV1-CMV_fwd
     primer_bind     63..84
                     /label=oEP143
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(238..257)
                     /label=oSEQ065 CMV_rev
     promoter        371..595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     528..548
                     /label=oSEQ030 CMV_fwd
     primer_bind     complement(543..562)
                     /label=oSEQ042 SFCp_rev
     primer_bind     complement(556..594)
                     /label=pPV1-CMV_rev
     primer_bind     complement(560..579)
                     /label=oEP138
     primer_bind     complement(560..579)
                     /label=oEP144
     primer_bind     575..592
                     /label=pPV1-EF1a-bb_fwd
     primer_bind     complement(576..635)
                     /label=oEP204
     primer_bind     complement(576..601)
                     /label=oEP202
     primer_bind     576..595
                     /label=oEP206 CMV-rev
     misc_feature    596..658
                     /label=hPGK 5' UTR
     primer_bind     619..678
                     /label=oEP203
     primer_bind     655..678
                     /label=oEP201
     regulatory      663..672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     666..687
                     /label=GG_mRuby2_fwd
     primer_bind     668..687
                     /label=mRuby2_fwd
                     /note="Fwd primer for mRuby2"
     CDS             669..1379
                     /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     669..688
                     /label=oEP159
     primer_bind     complement(695..716)
                     /label=oEP076
     primer_bind     717..736
                     /label=oEP073
     primer_bind     739..758
                     /label=oSEQ062 mRuby2_fwd
     primer_bind     complement(739..758)
                     /label=oSEQ093 mRuby2_rev
     primer_bind     complement(1271..1290)
                     /label=oSEQ061 mRuby2_rev
     primer_bind     1305..1324
                     /label=OSEQ091 mRuby2_fwd_SEQ
     primer_bind     complement(1360..1380)
                     /label=oEP160
     primer_bind     complement(1364..1391)
                     /label=GG_mRuby2_rev
     primer_bind     complement(1364..1386)
                     /label=gemRuby2_rev
     primer_bind     complement(1364..1383)
                     /label=mRuby2_rev
                     /note="Rev primer for mRuby2"
     primer_bind     1382..1406
                     /label=gebGH_fwd
     primer_bind     1388..1409
                     /label=GG_bGH_fwd
     primer_bind     1392..1409
                     /label=GG_WPRE-bGH_fwd
     polyA_signal    1394..1618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1478..1503)
                     /label=oEP181 IVT-bGH-rev
     misc_feature    1501..1503
                     /label=bGH TES
     primer_bind     complement(1600..1622)
                     /label=gebGH_rev
     primer_bind     complement(1601..1617)
                     /label=oSEQ078 bGH_rev
     primer_bind     complement(1604..1618)
                     /label=GG_bGH_rev
     primer_bind     1619..1660
                     /label=oMJ001
     primer_bind     1619..1660
                     /label=oMJ005
     primer_bind     1621..1660
                     /label=oMJ009
     primer_bind     1621..1660
                     /label=oMJ013
     primer_bind     1627..1657
                     /label=oMJ020
     primer_bind     complement(1639..1664)
                     /label=oSEQ033 Pentr_rev
     misc_feature    1641..1664
                     /label=pENTR2B.Seq.REV
     primer_bind     complement(1653..1674)
                     /label=oTA069 mIR-backbone_junction_rev
     primer_bind     complement(1653..1674)
                     /label=oSEQ044 pENTR_rev_seq
     primer_bind     complement(1726..1746)
                     /label=oSEQ005 pENTR_rev
                     /note="25nm	STD"
     CDS             1740..2549
                     /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(1765..1786)
                     /label=oSEQ023 attl2_rev
     primer_bind     complement(1773..1796)
                     /label=Kan_rev_SEQ
     primer_bind     complement(2163..2192)
                     /label=oTA099 GG_entry_upstream_fwd
     primer_bind     2163..2192
                     /label=oTA102 GG_entry_downstream_rev
     misc_feature    2623..3242
                     /label=pBR322_Origin
     rep_origin      2639..3227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2671..2690
                     /label=oEP033
     primer_bind     complement(2671..2690)
                     /label=oEP035
     primer_bind     2991..3011
                     /label=oEP024
     primer_bind     complement(2991..3011)
                     /label=oEP025
     primer_bind     3034..3049
                     /label=oTA073 Xho_addition_rev
     primer_bind     complement(3104..3119)
                     /label=oTA072 Xho_addition_for
     primer_bind     3128..3147
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     complement(3244..3263)
                     /label=oEP013
     primer_bind     3244..3263
                     /label=oEP014
     primer_bind     3379..3392
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      3392..3478
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3541..3560
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      3570..3597
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
       61 cttactcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctgagctc gtttagtgaa ccgtcctccg
      601 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagaaa
      661 tggccaccat ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg
      721 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc
      781 cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg
      841 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga
      901 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta
      961 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct
     1021 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc
     1081 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc
     1141 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt
     1201 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg
     1261 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac
     1321 gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt
     1381 aacaacgttg cacctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1441 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1501 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1561 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg
     1621 ctcgctgact gcaggtgccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1681 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1741 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1801 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1861 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1921 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1981 ccatcaagca ttttatccgt actcctggtg atgcatggtt actcaccact gcgatccccg
     2041 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2101 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2161 gcgatcgcgt atttcgtcta gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2221 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2281 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2341 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2401 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2461 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     2521 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacattatt
     2581 cagattgggc cccgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2701 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3301 tatcccctga ttctgtggat aaccgtatta ccgctagcat ggatctcggg gacgtctaac
     3361 tactaagcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga
     3421 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3481 gccgggagcg gatttgaacg ttgtgaagca acggcccgga gggtggcggg caggacgccc
     3541 gccataaact gccaggcatc aaactaagca gaaggccatc ctgacggatg gccttttt
//
