LOCUS       sequence-12833-e        6020 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 6020)
  AUTHORS   Fusco D, Accornero N, Lavoie B, Shenoy SM, Blanchard JM, Singer RH, 
            Bertrand E
  TITLE     Single mRNA molecules demonstrate probabilistic movement in living 
            mammalian cells.
  JOURNAL   Curr Biol. 2003 Jan 21. 13(2):161-7.
  PUBMED    12546792
REFERENCE   2  (bases 1 to 6020)
  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: "Curr Biol. 
            2003 Jan 21. 13(2):161-7."
COMMENT     Sequence Label: pMS2-YFP
FEATURES             Location/Qualifiers
     source          1..6020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             750..776
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             750..776
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             777..788
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1173..1182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1179..1181,1182..1184,1185..1895)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1224..1245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1485..1504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1832..1853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1896..1916
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1975..2547
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2679..2698)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2744..2799
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2798..2817)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3153..3172)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3154..3172)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3179..3196)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3179..3195)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3188..3210)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3337..3792
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3424..3443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3634..3655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3818..3922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4107..4486
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(4141..4160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4955..5543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5444..5463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5697..5714
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5831..5852
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5867..5872,5873..5890,5891..5897)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5905..5921
                     /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)."
     primer_bind     5910..5932
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5929..5945
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5929..5945
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5966..5984
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tctagagtcg acctgcagcc caagcttggg ctgaacatcg agcgccaggg ctccgtaaag
       61 ctactagagc acaggcggtg ccccaacgtc ctggggcctc tccactaata acggctactt
      121 ccaattgatt ggacgcgcca tcttgcctgc cttatgcata ttcagcggtg aactgaatat
      181 tcatgaacga ggcccgtccc gtccctccct cccttccccc ccccccggaa cccgctccgg
      241 aggacccgaa gggccccgcc ttcattaccg atgcgtagga caaaccattt tcccgatgtg
      301 tgtgggggga tactaatgag agactttagc tgaaaatgag cctgaactcc gaagctgagt
      361 aaaaatggcc taactttatc ctccgttctg taagtcctcg gtttgagtgc acgggaaacc
      421 cgaaaggagg acgacaggac caggaacatt ctcctcctcc tgtcgcgtca gaaagaacac
      481 ccaaccaggg agccggagcc ctagcgtcaa caactccgcc gcgcgcgctc cgtgtaggcc
      541 ggtgcgggcg gccccgtagc gcaagggagg gcgggaaagg aaggggcggg acacaagggc
      601 gaatctataa agggcgtcac tcagccagtt ctctcctcag aagcgccgag agcgcgaccg
      661 ggacggttgg agaagaaggt ggctcccgga agggggagag acaaactgcc gtaacctctg
      721 ccgttcagga tcactagtcc accatgggct acccctacga cgtgcccgac tacgccatcg
      781 aaggccgcca tatgctagcc gttaaaatgg cttctaactt tactcagttc gttctcgtcg
      841 acaatggcgg aactggcgac gtgactgtcg ccccaagcaa cttcgctaac gggatcgctg
      901 aatggatcag ctctaactcg cgttcacagg cttacaaagt aacctgtagc gttcgtcaga
      961 gctctgcgca gaatcgcaaa tacaccatca aagtcgaggt gcctaaaggc gcctggcgtt
     1021 cgtacttaaa tatggaacta accattccaa ttttcgccac gaattccgac tgcgagctta
     1081 ttgttaaggc aatgcaaggt ctcctaaaag atggaaaccc gattccctca gcaatcgcag
     1141 caaactccgg catctacgcg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1201 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1261 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1321 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     1381 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1441 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1501 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1561 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1621 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1681 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1741 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
     1801 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1861 ccgggatcac tctcggcatg gacgagctgt acaagccaaa aaagaagaga aaggtataaa
     1921 tcgattctgc agtagctcgc cggcagctga ctagctagag ctgagaactt cagggtgagt
     1981 ttggggaccc ttgattgttc tttctttttc gctattgtaa aattcatgtt atatggaggg
     2041 ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc ccttgtatca ccatggaccc
     2101 tcatgataat tttgtttctt tcactttcta ctctgttgac aaccattgtc tcctcttatt
     2161 ttcttttcat tttctgtaac tttttcgtta aactttagct tgcatttgta acgaattttt
     2221 aaattcactt ttgtttattt gtcagattgt aagtactttc tctaatcact tttttttcaa
     2281 ggcaatcagg gtatattata ttgtacttca gcacagtttt agagaacaat tgttataatt
     2341 aaatgataag gtagaatatt tctgcatata aattctggct ggcgtggaaa tattcttatt
     2401 ggtagaaaca actacaccct ggtcatcatc ctgcctttct ctttatggtt acaatgatat
     2461 acactgtttg agatgaggat aaaatactct gagtccaaac cgggcccctc tgctaaccat
     2521 gttcatgcct tcttctcttt cctacagctc ctgggcaacg tgctggttgt tgtgctgtct
     2581 catcattttg gcaaagaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     2641 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     2701 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     2761 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     2821 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     2881 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     2941 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     3001 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     3061 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     3121 agatccctcg agcggccgcg gtacccaatt cgccctatag tgagtcgtat tacaattcac
     3181 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3241 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3301 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3361 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3421 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3481 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3541 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3601 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3661 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3721 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3781 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4141 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaaaca
     4381 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4441 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4501 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4561 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4621 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4681 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4741 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5641 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5701 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5761 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5821 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5881 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5941 atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggagc
     6001 tccaccgcgg tggcggccgc
//
