LOCUS       pKG3831_pGEEC579        3406 bp    DNA     circular SYN 27-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3406)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3406
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     complement(14..58)
                     /label=oMJ002
     primer_bind     complement(14..54)
                     /label=oMJ010
     primer_bind     complement(14..50)
                     /label=oMJ006
     primer_bind     complement(14..50)
                     /label=oMJ014
     primer_bind     complement(22..50)
                     /label=oMJ019
     primer_bind     55..76
                     /label=pPV1-UbC_fwd
     promoter        59..458
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(191..210)
                     /label=oSEQ007
     primer_bind     complement(191..210)
                     /label=oSEQ007 UBC_rev
     primer_bind     401..415
                     /label=oEP187 IVT-UbC-fwd
     misc_feature    401..406
                     /label=UbC TSS
     primer_bind     436..455
                     /label=oEP208 UbC-rev
     primer_bind     437..476
                     /label=oEP185
     primer_bind     complement(437..476)
                     /label=oEP186
     misc_feature    456..457
                     /label=INTRON REMOVAL SITE
     regulatory      463..472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     466..487
                     /label=GG_mRuby2_fwd
     primer_bind     468..487
                     /label=mRuby2_fwd
                     /note="Fwd primer for mRuby2"
     CDS             469..1179
                     /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     complement(495..516)
                     /label=oEP076
     primer_bind     517..536
                     /label=oEP073
     primer_bind     539..558
                     /label=oSEQ062 mRuby2_fwd
     primer_bind     complement(539..558)
                     /label=oSEQ093 mRuby2_rev
     primer_bind     complement(1071..1090)
                     /label=oSEQ061 mRuby2_rev
     primer_bind     1105..1124
                     /label=OSEQ091 mRuby2_fwd_SEQ
     primer_bind     complement(1164..1191)
                     /label=GG_mRuby2_rev
     primer_bind     complement(1164..1186)
                     /label=gemRuby2_rev
     primer_bind     complement(1164..1183)
                     /label=mRuby2_rev
                     /note="Rev primer for mRuby2"
     primer_bind     1182..1206
                     /label=gebGH_fwd
     primer_bind     1188..1209
                     /label=GG_bGH_fwd
     primer_bind     1192..1209
                     /label=GG_WPRE-bGH_fwd
     polyA_signal    1194..1418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1278..1303)
                     /label=oEP181 IVT-bGH-rev
     misc_feature    1301..1303
                     /label=bGH TES
     primer_bind     complement(1400..1422)
                     /label=gebGH_rev
     primer_bind     complement(1401..1417)
                     /label=oSEQ078 bGH_rev
     primer_bind     complement(1404..1418)
                     /label=GG_bGH_rev
     primer_bind     1419..1460
                     /label=oMJ001
     primer_bind     1419..1460
                     /label=oMJ005
     primer_bind     1421..1460
                     /label=oMJ009
     primer_bind     1421..1460
                     /label=oMJ013
     primer_bind     1427..1457
                     /label=oMJ020
     primer_bind     complement(1439..1464)
                     /label=oSEQ033 Pentr_rev
     misc_feature    1441..1464
                     /label=pENTR2B.Seq.REV
     primer_bind     complement(1453..1474)
                     /label=oTA069 mIR-backbone_junction_rev
     primer_bind     complement(1453..1474)
                     /label=oSEQ044 pENTR_rev_seq
     primer_bind     complement(1526..1546)
                     /label=oSEQ005 pENTR_rev
                     /note="25nm	STD"
     CDS             1540..2349
                     /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(1565..1586)
                     /label=oSEQ023 attl2_rev
     primer_bind     complement(1573..1596)
                     /label=Kan_rev_SEQ
     primer_bind     complement(1963..1992)
                     /label=oTA099 GG_entry_upstream_fwd
     primer_bind     1963..1992
                     /label=oTA102 GG_entry_downstream_rev
     misc_feature    2423..3042
                     /label=pBR322_Origin
     rep_origin      2439..3027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2834..2849
                     /label=oTA073 Xho_addition_rev
     primer_bind     complement(2904..2919)
                     /label=oTA072 Xho_addition_for
     primer_bind     2928..2947
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     3179..3192
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      3192..3278
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3341..3360
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      3370..3397
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     join(3399..3406,1..12)
                     /label=pENTR_fwd_seq
     primer_bind     join(3399..3406,1..12)
                     /label=oSEQ043 pENTR_fwd_seq
ORIGIN
        1 acaaactctt cctgttagtt agttacttaa gctcgggccc acctgcagtc tacttactgg
       61 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc
      121 acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg
      181 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
      241 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
      301 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
      361 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
      421 cggttcttgt ttgtggatcg ctgtgatcgt cacttgacaa tggccaccat ggtgtctaag
      481 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
      541 ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg aactcaaacc
      601 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
      661 tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc tgatttcttt
      721 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
      781 gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta ccacgtccaa
      841 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
      901 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactcatatg
      961 gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac ttacaggtca
     1021 aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca ccgcctggaa
     1081 aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc agttgccaag
     1141 ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt aacaacgttg cacctgtgcc
     1201 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1261 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1321 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1381 caatagcagg catgctgggg atgcggtggg ctctatggcg ctcgctgact gcaggtgccc
     1441 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1501 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1561 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1621 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     1681 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1741 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1801 actcctggtg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     1861 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1921 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtcta
     1981 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2041 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2101 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2161 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2221 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2281 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2341 tttttctaat cagaattggt taattggttg taacattatt cagattgggc cccgttccac
     2401 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2461 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2521 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2581 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2641 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2701 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2761 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2821 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2881 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2941 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3001 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3061 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3121 aaccgtatta ccgctagcat ggatctcggg gacgtctaac tactaagcga gagtagggaa
     3181 ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga ctgggccttt cgttttatct
     3241 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3301 ttgtgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3361 aaactaagca gaaggccatc ctgacggatg gcctttttgc gtttct
//
