LOCUS       sequence-219911-        6362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6362)
  AUTHORS   Miller AL, Bement WM
  TITLE     Regulation of cytokinesis by Rho GTPase flux.
  JOURNAL   Nat Cell Biol. 2009 Jan . 11(1):71-7.
  PUBMED    19060892
REFERENCE   2  (bases 1 to 6362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2009 Jan . 11(1):71-7."
COMMENT     Sequence Label: EMTB-mCherry
FEATURES             Location/Qualifiers
     source          1..6362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..1024
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1059..1077
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1059..1076
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1109..1118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1115..1822
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1829..2536
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             2561..2590
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3438..3572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3462..3481)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3516..3535
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3685..3705)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3685..3703)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3724..3740)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3724..3740)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3737..3759)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3748..3764
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3772..3778,3779..3796,3797..3802))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3817..3838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3955..3972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4126..4714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4206..4225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4885..5676,5677..5745))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5508..5527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5746..5850)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5876..6331)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6013..6034)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6225..6244
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg
       61 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
      121 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
      181 gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg
      241 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
      301 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
      361 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
      421 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
      481 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
      541 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
      601 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
      661 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
      721 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
      781 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
      841 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
      901 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
      961 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
     1021 gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca ctatagaata
     1081 caagctactt gttctttttg caggatccgc caccatggtg agcaagggcg aggaggataa
     1141 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     1201 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1261 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     1321 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     1381 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     1441 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     1501 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     1561 ggcctcctcc gagagaatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     1621 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     1681 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     1741 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     1801 cggcatggac gagctgtaca agtccagaat ggtgagcaag ggcgaggagg ataacatggc
     1861 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     1921 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1981 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     2041 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2101 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2161 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2221 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2281 ctccgagaga atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2341 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2401 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     2461 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     2521 ggacgagctg tacaagtccg gactcagatc tcgagatatg gagcagaagc tcatctcaga
     2581 agaagacctc ggcgcgccag tgcgaagcga aacagcaccc gacagctaca aagtgcaaga
     2641 taagaaaaat gcctccagcc gccctgcctc tgcaatttca ggacaaaata acaaccactc
     2701 aggaaataaa ccagaccctc cgcctgtgtt acgtgttgat gaccggcagc ggctggcccg
     2761 ggagcgacgt gaggaacggg agaaacagct agctgcaaga gaaatagtgt ggttagaaag
     2821 agaagagcga gccaggcagc actacgagaa gcacctggaa gagcggaaga agaggttgga
     2881 ggagcagagg cagaaggagg agcggaggag ggctgctgtg gaggagaagc ggaggcagag
     2941 acttgaggag gacaaagaac gccacgaagc tgttgtacgg cgcacaatgg aaaggagcca
     3001 gaagccaaaa cagaagcata accgttggtc gtggggaggc tctctccatg ggagccctag
     3061 catccacagt gcagatccag acaggcggtc agtttccacc atgaatcttt cgaaatatgt
     3121 tgatcccgtc attagcaagc ggctctcctc ttcatctgca actttactaa attctccaga
     3181 tagagctcgc cgcctgcagc tcagcccatg ggagagcagc gttgttaaca gactcctgac
     3241 gcccacacat tcgttcctgg ccagaagtaa aagcacagct gccttgtctg gagaagcagc
     3301 atcttgcagc cccatcatca tgccctacaa agctgcacac tctagaaatt cgatggatcg
     3361 accaaaactc tttgtaacac cacctgaggg ctcttaggaa ttctgcagtc tagaactata
     3421 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     3481 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3541 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3601 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     3661 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3721 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3781 agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3841 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5881 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5941 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6001 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6061 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6121 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6181 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6241 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6301 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     6361 tg
//
