LOCUS       40924_1554        7835 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector L232-3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7835)
  AUTHORS   Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, 
            Klimovich BV, Samoilovich MP, Satou Y, Kruss M, Gelhaus C, Kurn U, 
            Bosch TC, Khalturin K.
  TITLE     Blood System Formation in the Urochordate Ciona intestinalis 
            Requires the Variable Receptor vCRL1
  JOURNAL   Mol. Biol. Evol. 29 (10), 3081-3093 (2012)
  PUBMED    22513285
REFERENCE   2  (bases 1 to 7835)
  AUTHORS   Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, 
            Klimovich VB, Samoilovich MP, Satou Y, Kruess M, Gelhaus C, Kuern U,
            Bosch TCG., Khalturin K.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-FEB-2011) Zoological Institute, Christian-Albrechts 
            University Kiel, Am Botanischen Garten 9, Kiel, Schleswig-Holstein 
            24118, Germany
REFERENCE   3  (bases 1 to 7835)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7835)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Biol. 
            Evol."; date: "2012"; volume: "29"; issue: "10"; pages: "3081-3093"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-FEB-2011) Zoological Institute, Christian-Albrechts University 
            Kiel, Am Botanischen Garten 9, Kiel, Schleswig-Holstein 24118, 
            Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2448..3161
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        complement(4992..5010)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5020..5036)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5178..5633
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5659..5763
                     /label=AmpR promoter
     CDS             5764..6621
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6795..7383
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7671..7692
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7707..7737
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7745..7761
                     /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     7769..7785
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7806..7824
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtaccaagt gcttgaaatg ctaaatgttt tcaatttttc gccattaaga caagcctaca
       61 caaatgcttc tataaattat gccaagcacg ttagcagctt ctacgagccc caaccactat
      121 taattcgaac agcatgtttt tttgcagtgc gcaatgttta acacactata ttatcaatac
      181 tactaaagat aacacatacc aatgcatttc gtctcaaaga gaattttatt ctcttcacga
      241 cgaaaaaaaa agttttgctc tatttccaac aacaacaaaa atatgagtaa tttattcaaa
      301 cggtttgctt aagagataag aaaaaagtga ccactattaa ttcgaacgcg gcgtaagctt
      361 accttaatct caagaagagc aaaacaaaag caactaatgt aacggaatca ttatctagtt
      421 atgatctgca aataatgctg cagggaatgt aacttgcttt accatcgcgt ggtgggaaaa
      481 cgacagtcgt tatagcacga gtgttcatac acctcgtgcc agcttacgag ctaccatata
      541 tgttgtgggc gaataaaggt tttataaata taacaggttt tataaatata acaacgccat
      601 tttaaagtcg gttacataat tctgtaacta gttcaaattg aacggtaaac gtaaataaaa
      661 accttgaccg tcttacccaa ttatataaaa acactttgaa cgctttttag gatggaaggg
      721 tatggccatg ctagataatt ctgtggacca tctcaaccca acctattaca gaacggtcgt
      781 aataatgaaa atggatacca tttttaggca tatagactga ttcctctact ttctagaaac
      841 gtaagcagta tacacagaaa aaatgaagtg tgattctgtg caattaaacc gttctaaatt
      901 catagccgac tgaatttcta attaagtgaa tgtctgacct agatttattg ttaagtttag
      961 caccaaatct gagccagcga taagcagtct aattaaattg gctgctggcg ataaaatagg
     1021 tcatcctgaa aaatcgtttg cgcctttatt taaaatatag tagagtgggg aaagacggga
     1081 catcttatcg ttctattttc tcgtcccatt tcgtagtaaa caaagaacat tcaaaaaata
     1141 taaaaccata acttcaaaac ttcaatagac cgttgtcaac tgtttaaaac acagtaagag
     1201 aatttggata ttatgtgcta aaggtgtccc atctcccccc accctactat atctgtttat
     1261 agttctgtgg ggtaagatga gataccgtta acacctaaac atttttactt taaacaatca
     1321 accacgtttt ttatggtcgt gatgggcatg aggttacata attctgaaaa tattttttgc
     1381 ccccgaccaa aagacgcgaa gagtaaaaac atgtctcagc ttatattccc cacataaata
     1441 tatttttgta ctgtttggtg aatttataaa cttatattac catgcatata cgttatgtta
     1501 ctggtatttt ctcagtaggc aaattcattt gtccacgttt tataggtttt tatttatgat
     1561 ttttaaaatg ctaaaaatgt gggagggggt tgaaagtaca atacaaacac acaaaacaac
     1621 tcaaactaaa gatttatagt tatgctaatt cacctacaca atataacaag atgtgtaatg
     1681 caaccatgtg tttatgatga gcgctaacat attttgtaac cactcaaatt ccccgccaca
     1741 cgaggataat gaataggtga ctctgtagtc tgtacatctt agactgaaat aaagattata
     1801 aatctacgaa ataaaataat ttctgctcac tgattatact tctgttttat agattagaaa
     1861 ccgtttctaa taaatgacct aattcgctat acacacacgc tgtgcgcgag ataatcattc
     1921 tcgcaccccg tttattgtgt taaaattgcc gcctagattc acaaagcgtg acggctagag
     1981 ccagcaacgt gtcgccttca attacgcaac atccgggttg cgcaattctg gatataaaag
     2041 aactaacaaa gatgacgtag ctaccttttt cagttcagac ttacgaaaga ctcacgtgtc
     2101 ggcggtctac ttgtcctttt cgagctgtgg caatttggtg agtggttcta tcttatatct
     2161 gagtacatct ctaaggaatt atagtttgat tagttaagtt tttattgtta ggaaagatga
     2221 aatcattagg ttttacttag tttaagtatg ttagtactgg ttaggcgttt gaattattga
     2281 aaaactcagt tcgttaactg tagtagttct ggtagcttag caagtatacc ctgtatacgc
     2341 cttttggctt tttaacaata acttaaactt attttacagc aaatttctgt gcattcggtt
     2401 aaccccaacc ttccaaaatg aaagacaaga ctttaattaa tgaaaaaatg agtaaaggag
     2461 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     2521 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     2581 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttct
     2641 gttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat gactttttca
     2701 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     2761 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     2821 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     2881 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact
     2941 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     3001 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
     3061 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     3121 cagctgctgg gattacacat ggcatggatg aactatacaa agcgatcgcg tgactgcagc
     3181 caacgacacc gcccaccatc acttctgcag ttttactcca ccagcttgct cacccttcta
     3241 accatttcag cctagttaaa cccccagttc aatttgtcag ccttttaaat tgcgtcatga
     3301 catttcagtt gttactgtgt gccgctctgt ccacatggtg tagttatgtt tttcattacc
     3361 cccaccgaaa acactttcgt aggaaaaaac cctccgtttc gtgccaactt aaaagtgtta
     3421 tttttgtttc tagcgctgct taacttattt aaaatgacaa gactatctat gattttaaac
     3481 catggctttg aatggcgtac atgtattatt gtgtttatat tcagttaaac cgtactatta
     3541 ctgcttggaa aattcagttc attccttcac cgaaggcttt aacgtggttg gttgcggtgt
     3601 tgtcaatttg ctgctggaaa taaagcttct gttaagtgta tactgtctct tggtttgcaa
     3661 gattattatg gatccagatg ttgtatatac aagagtgaag atttagcgtt tttatgtggt
     3721 aaaacgccca tggtgatcta tttacagttt ttaagttgag ctgatcaatt ttcacatgat
     3781 gatcctgaac gaccaggggc acaaaggcat ttgcagattg tcccatctct cactacgacc
     3841 gatctaccac tattttcaca acggccagta catttcgaag tctggtattt agcgaagtag
     3901 tgttcaattg ccaaagccat gtttctctgc ttcgctctgt ctgggatgac cgatgatatg
     3961 ggtcgcagct gtaaaattac aaatagtcag tttttctgac gaaagtgaat gttgattaca
     4021 ttcaaatgaa agtcgttgat ggctaatgcc atcgttcata cgctttaatt agcagattat
     4081 gtggttatat tagatatctt taccagtaat attgagaacg ccaaggtaaa atctagtttt
     4141 ttgatactta atgaacagcc atattaaaag ccatacatga aaaaaaaatt aaaaaaaaca
     4201 tacataaact ctcataagct gacacagtag gtcgtggctg acacgaggtg tatgaaacaa
     4261 aacgctcgtg ttttaacgac tgttgttgca ccgtcacgcg agaataaagt aacttccatt
     4321 catacatgat acagttctta ccttgaaatc gatgactgca gggctaagtt taactgatgc
     4381 tgtccagttt tcgtaagcat ttgttgtaag tcggtttcta gcaaagaaac tgagagcact
     4441 agctgtatga gactgcccac caatcacatt cgatataact tccttagcag ccattgtaaa
     4501 acttcctaaa tattagtaaa taacgaataa taatgtgtac atgactctgc ttatcagtta
     4561 gattgggcgc gccatcagtt taaacgaatg cggccgccac cgcgggacgt tacacaattc
     4621 taatattaat taaattattg ttttaagtat gatagtaaat cacattacgc cgcgttcgaa
     4681 ttaatagtgg tcactttttt cttatctctt aagcaaaccg tttgaataaa ttactcatat
     4741 ttttgttgtt gttggaaata gagcaaaact ttttttttcg tcgtgaagag aataaaattc
     4801 tctttgagac gaaatgcatt ggtatgtgtt atctttagta gtattgataa tatagtgtgt
     4861 taaacattgc gcactgcaaa aaaaacatgc tgttcgaatt aatagtggtt ggggctcgta
     4921 gaaaacgaaa aatatcttaa gctagcatag agaatggagc aaaactcaat ttgatgcgag
     4981 ctccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     5041 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5101 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5161 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5221 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5281 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5341 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5401 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5461 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5521 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5581 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5641 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5701 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5761 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5821 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5881 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5941 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6001 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6061 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6121 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6181 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6241 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6301 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6361 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6421 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6481 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6541 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6601 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6661 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6721 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6781 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6841 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6901 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6961 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7021 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7081 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7141 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7201 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7261 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7321 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7381 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7441 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7501 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7561 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7621 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7681 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7741 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7801 cgcgcaatta accctcacta aagggaacaa aagct
//
