LOCUS       Exported                6019 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMS2-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6019)
  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 6019)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6019
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1179..1895
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1179..1895
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     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             3923..4783
                     /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     complement(4141..4160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4954..5542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5443..5462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5696..5713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5830..5851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5866..5896
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5904..5920
                     /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     5909..5931
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5928..5944
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5928..5944
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5965..5983
                     /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 ccttcccccc acccccggaa 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 ctgacctacg
     1381 gcgtgcagtg cttcagccgc 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 cctgagcacc 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 tgcacaacat
     4381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4441 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5701 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5761 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5821 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5881 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     5941 tgaccatgat tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggagct
     6001 ccaccgcggt ggcggccgc
//
