LOCUS       40924_44524        5514 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTVP1GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5514)
  AUTHORS   Garcia-Fruitos E, Gonzalez-Montalban N, Morell M, Vera A, Ferraz RM,
            Aris A, Ventura S, Villaverde A.
  TITLE     Aggregation as bacterial inclusion bodies does not imply 
            inactivation of enzymes and fluorescent proteins
  JOURNAL   Microb. Cell Fact. 4, 27 (2005)
  PUBMED    16156893
REFERENCE   2  (bases 1 to 5514)
  AUTHORS   Garcia-Fruitos E.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2014) Institut de Biotecnologia i de Biomedicina, 
            CIBER-BBN, Autonomous University of Barcelona, Campus Bellaterra, 
            Bellaterra 08193, Spain
REFERENCE   3  (bases 1 to 5514)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5514)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact. 4, 27 (2005)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JUL-2014) Institut de Biotecnologia i de Biomedicina, CIBER-BBN,
            Autonomous University of Barcelona, Campus Bellaterra, Bellaterra 
            08193, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     CDS             909..1619
                     /codon_start=1
                     /label=GFPuv
                     /note="GFP variant optimized for excitation by UV light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      1867..1953
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2045..2072
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2092..2183
                     /label=AmpR promoter
     CDS             2184..3041
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3215..3803
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3989..4129)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        4315..4392
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4393..5472
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg aaactacgac cactggtgaa tctgctgacc
      301 ccgtcaccac taccgttgag aactacggag gagagactca ggtccaacgt cgccaccaca
      361 ccgacgttgc cttcgttctt gaccggtttg tggaggtcac agtgtcggat aaccaacaca
      421 cactcgacgt gatgcaggca cacaaagaca atatcgtggg cgcgcttctt cgcgcagcca
      481 cgtactattt ttctgatttg gaaatagcag tgacccacac tgggaagctc acatgggtgc
      541 ccaacggtgc accagtttct gcacttaaca acacaaccaa tcccactgcc taccacaagg
      601 gcccggtgac tcgactggct ctcccataca ccgcgccaca ccgtgtgttg gctacggcgt
      661 acactggcac tacgacctac accgccagtg cacgcgggga ttcggctcac ctaacgacga
      721 cgcatgctcg gcatttgccg acatcgttca actttggtgc agttaaagca gaaacaatca
      781 ctgagttgct cgtgcgcatg aagcgtgctg aactctattg tcctaggccg attcttccga
      841 ttcagccaac gggcgataga cgcaagcaac agctcgtcgc acctgcaaaa caactgctgg
      901 ggatcccgag caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
      961 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgctacat
     1021 acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa
     1081 cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga
     1141 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat
     1201 ctttcaaaga tgacgggaac tacaagacgc gtgctgaagt caagtttgaa ggtgataccc
     1261 ttgttaatcg tatcgagtta aaaggtattg attttaaaga agatggaaac attctcggac
     1321 acaaactcga gtacaactat aactcacaca atgtatacat cacggcagac aaacaaaaga
     1381 atggaatcaa agctaacttc aaaattcgcc acaacattga agatggttcc gttcaactag
     1441 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
     1501 attacctgtc gacacaatct gccctttcga aagatcccaa cgaaaagcgt gaccacatgg
     1561 tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat
     1621 aaggaattct gagtaggtaa gtcgacctgc aggcatgcaa gcttggctgt tttggcggat
     1681 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     1741 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     1801 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     1861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1921 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     1981 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     2041 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct ttttgtttat
     2101 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2161 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2221 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2281 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2341 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2401 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2461 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2521 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2581 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2641 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2701 acgacgagcg tgacaccacg atgcctacag caatggcaac aacgttgcgc aaactattaa
     2761 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2821 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2881 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2941 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3001 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3061 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3121 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3181 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3241 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3301 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3361 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3421 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3481 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3541 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3601 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3661 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3721 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3781 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3841 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3901 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3961 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4021 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4081 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     4141 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4201 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4261 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc
     4321 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4381 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4441 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4501 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4561 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4621 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4681 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4741 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4801 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     4861 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     4921 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     4981 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     5041 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     5101 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     5161 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     5221 gaagacagct catgttatat cccgccgtca accaccatca aacaggattt tcgcctgctg
     5281 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     5341 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     5401 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5461 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag cgcgaattga tctg
//
