LOCUS       62056_1291        5639 bp DNA     circular SYN 23-JAN-2019
DEFINITION  Expression vector Hym-176E_EGFP DNA, complete sequence.
ACCESSION   LC426376
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5639)
  AUTHORS   Noro Y, Yum S, Nishimiya-Fujisawa C, Busse C, Shimizu H, Mineta K, 
            Zhang X, Holstein TW, David CN, Gojobori T, Fujisawa T.
  TITLE     Regionalized nervous system in Hydra and the mechanism of its 
            development
  JOURNAL   Gene Expr. Patterns (2019) In press
REFERENCE   2  (bases 1 to 5639)
  AUTHORS   Noro Y, Fujisawa T, Shimizu H.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-SEP-2018) Contact:Yukihiko Noro Waseda University, 
            Center for Advanced Biomedical Sciences (TWIns); 2-2 Wakamatsu-cho, 
            Shinjuku-ku, Tokyo 162-8480, Japan
REFERENCE   3  (bases 1 to 5639)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene Expr. 
            Patterns (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-SEP-2018) Contact:Yukihiko Noro Waseda University, Center for 
            Advanced Biomedical Sciences (TWIns); 2-2 Wakamatsu-cho, 
            Shinjuku-ku, Tokyo 162-8480, Japan"
FEATURES             Location/Qualifiers
     source          1..5639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        362..466
                     /label=AmpR promoter
     CDS             467..1324
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1498..2086
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2374..2395
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2410..2440
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2448..2464
                     /label=lac operator
                     /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     2472..2488
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4399..5112
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
       61 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
      121 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
      181 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
      241 gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata
      301 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
      361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      481 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      541 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      601 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      661 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      721 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      781 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      841 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
      901 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
      961 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1021 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1081 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1141 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1201 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1261 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1321 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1381 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1861 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     1921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2281 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2341 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     2401 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2461 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcagct
     2521 tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc gcccttgctt
     2581 agtgattaga agtattccct ctgttgaaaa attttttatc ataaaaacaa atttgccatt
     2641 attatatgct aaccaatcca cattggaatc ttctattagt tcttactgct ttctttgttg
     2701 cctttgttga ttatgttttg acgtaagtgc gaactaaagt ttttacacgc actgtcaggt
     2761 ggcttaagac agcagcttat aagaaatgct tttatttata ttgaagaatt gattataaca
     2821 atctgtttat tacggtcaga aacgctcaaa tcgtttctaa caaatgcata acttattttt
     2881 tacgtaaatc agaactcctc caccttgttt tttattatgt cttaataaca aaataatttt
     2941 caagtcagca atatgaaaat taattttatt taaaggccca cttaaataac accgaattta
     3001 tgttagttaa ataatgtttt aaggtacttt taaggtagta ttaaaaagtc aatcaataac
     3061 attttgtgtt gagaatttaa aggtgagtta tcactcatcc gtttttctac gggaacagga
     3121 tagggaaggg aaaaataatc acgtgggtat acgtattata agttttaatt tgtccaaagg
     3181 aaaactgcgc atgatcacgt gattatttta tccgttcccg caaaaaaacg gaagagtgga
     3241 aactcacctc aaagcggact attgggataa cattttatga cgtcatttaa tagttacaaa
     3301 ggtaaagaag tataagacta atattcaaaa aaaaattaag taaaatagtt ttatgatttg
     3361 ttgctgaaaa tatataaaaa ttgcttacct cccccccccc ctatttcaaa aaaaatctta
     3421 gtttaaaaaa aaacaacgtt aaaaacatag accagataat tggttaaaaa ttaatttatc
     3481 tcaaatcagt tgaaaataaa aatctataaa ataggtcgaa aaacaaatcg gtcgaaaacg
     3541 aaaacgtcat aaatcgggtg aaaataaaaa aagaatatta tcgttcgaaa ataatttatg
     3601 tcttagtggg ttatttgaaa agtagttcca ctttttagta gatgttcatt ttttagaaca
     3661 acttagtagg gcaggatttt tgtaggtcta actatctgac acggctctac ccactacgcc
     3721 acagccgctt gataattaca tatgttgtaa taaaattaat cagttaaagt ctaattaatc
     3781 tcatcaaaat gaagaaacca acactaatct catcaacatg aaggaaccaa tactagcctc
     3841 atcaataaga agggacaaac gcttatctca taaacatgaa gaaattaaca ctaattgcat
     3901 caatacgaag gaaccagtat taatctcatc aatatgaaga aaccaacact aatctcatca
     3961 atatgaagaa accaacacta atctcatcaa aatgaaggaa ccagcgctaa tctcgtaaaa
     4021 atgaaggagc caactctaat ctcgtcagta tgaagtaagc aacattaatc tatttttcaa
     4081 gttgtaaaag actaaatcaa aaaatagcac gataaaaaaa tagttgttat gtagctagaa
     4141 agctagatca caccgggcat aggctatttt taatttaagt aacaaatgga aaattgattt
     4201 ttattaataa tgttctttct tgaaacaaca agttttattt tactataata aggtacaaaa
     4261 tcaatacaat agaattgatt gaaaaagttt ttatttaaca ttatttaacg cttattagat
     4321 aattgacttt ctcttaaact tattccgaaa cagtaagaag cagagaatct gatctcctct
     4381 aaagtacccg gggaaaaaat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
     4441 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     4501 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
     4561 gttccatggc caacacttgt cactactttc tgttatggtg ttcaatgctt ttcaagatac
     4621 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     4681 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
     4741 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     4801 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
     4861 gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga
     4921 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     4981 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     5041 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
     5101 gaactataca aatagcattc gtagaattca tcgtgatacg acaacgtaat aaaattttca
     5161 ttttttctga aaaaattaat gatttaattc ggtttttaat aattcaataa actatttatt
     5221 ttacaatttt ttgaacatgt tttattttgt tattgataat gcaaaaaaga tgttaaaata
     5281 ctttattgag ataaaaatag tttaaatgta atttgttgtc atttttatta attaattaaa
     5341 actaacataa acttttcaaa aacgcctagg acgtttttaa aacgtttaaa aaccttatta
     5401 aacgtttttt aaacgtttta aagacgttta tggcttgcta tgttccagtc gtagtttgaa
     5461 gctgctatgt ttttcaatgg taagacttaa tttgactttt ttttttgttt aagttccaat
     5521 aaattaatgt ccaggttgtt aaaatgttcg actgattaaa aatgctattt tttcaagata
     5581 tagacattat tctaaaaaat taaattttac tcttttttta aagagaaact ctcaccaac
//
