LOCUS       40924_2149        6411 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector mTol2-B-P-GFF-CG3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  AUTHORS   Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, 
            Chien CB, Dorsky R.
  TITLE     Large Scale Zebrafish Gal4 Enhancer Trap Screen for Neural Specific 
            Expression Patterns
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6411)
  AUTHORS   Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, 
            Chien CB, Dorsky R.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2013) Neurobiology and Anatomy, University of 
            Utah, 20 North 1900 East, Salt Lake City, UT 84132, USA
REFERENCE   3  (bases 1 to 6411)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-MAR-2013) Neurobiology and Anatomy, University of Utah, 20 North
            1900 East, Salt Lake City, UT 84132, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      181..636
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        714..818
                     /label=AmpR promoter
     CDS             819..1676
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1850..2438
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2726..2747
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2762..2792
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2800..2816
                     /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     2824..2840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2858..2876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    3144..3164
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     protein_bind    3248..3272
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3281..3721
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    complement(3822..3846)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4045..4065)
                     /label=attB3
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     primer_bind     complement(4073..4089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(4105..4239)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4360..4493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4544..5260)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(5261..6159)
                     /label=cmlc2
                     /note="Cardiac myosin light chain 2 promoter"
     promoter        complement(join(6410..6411,1..17))
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
       61 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      121 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      181 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      241 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      301 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      361 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      421 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      481 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      541 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      601 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
      661 gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat gtgcgcggaa
      721 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      781 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      841 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      901 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      961 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1021 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1081 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1141 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1201 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1261 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1321 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1381 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1441 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1501 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1561 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1621 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1681 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2521 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2581 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     2641 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2701 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     2761 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     2821 acacaggaaa cagctatgac catgattacg ccaagctatt taggtgacac tatagaatac
     2881 tcaagctatg catccaacgc gttgggagct ctcccatatg gtcgagcaga ggtgtaaaaa
     2941 gtactcaaaa attttactca agtgaaagta caagtactta gggaaaattt tactcaatta
     3001 aaagtaaaag tatctggcta gaatcttact tgagtaaaag taaaaaagta ctccattaaa
     3061 attgtacttg agtattaagg aagtagggat aacagggtaa tctcgagcag gaaacagcta
     3121 tgaccatgat tacgccaagc tatcaacttt gtatagaaaa gttgggccgg cccttcggcg
     3181 cgccactcta gagggtatat aatggatctc gcgtctcagc ctcactttga gctcctccac
     3241 accgcgccca agtttgtaca aaaaagcagg ctcggccacc atgaagctac tgtcttctat
     3301 cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa
     3361 gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc
     3421 tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct
     3481 atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca
     3541 ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt
     3601 cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat
     3661 aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc
     3721 gccggaattc ccggccgacg ccctggacga cttcgacctg gacatgctgc ctgctgatgc
     3781 tctcgatgat ttcgatctcg atatgctccc gggtaactaa gacccagctt tcttgtacaa
     3841 agtgggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3901 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3961 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4021 gggggaggtg tgggaggttt tttccaactt tattatacat agttgataat tcactggccg
     4081 tcgttttacg gtaccatcga tgatgatcca gacatgataa gatacattga tgagtttgga
     4141 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     4201 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     4261 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     4321 aaatgtggta tggctgatta tgatcctcta gatcagatct cttgtttatt gcagcttata
     4381 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4441 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctacgtaa
     4501 tacgactcac tatagttcta gaggctcgag aggggccgct ttacttgtac agctcgtcca
     4561 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     4621 cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca
     4681 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     4741 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     4801 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     4861 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     4921 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     4981 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     5041 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
     5101 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     5161 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     5221 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtcactgtc tgctttgctg
     5281 ttggtctggg ctcctgggtc actggcttac taatggagtc tttatgtatg aggactctta
     5341 tcaattgttc ttctataaag gtctgcagtg tttctgttcg tcccctacat ggacacccag
     5401 agcctcctaa atacaggagc cctgataact gcacaagtgc tcagattcca gcagggtgga
     5461 aaatgagata aagtgtgcag atggggaggg ggacgtgaat gagagatttg agggatgaaa
     5521 aggatggatg aacgcattga aaatagcccc tttcacaaaa taataccagt aaattgtcat
     5581 ataattaatg accagacctt tacctgtaaa tgatgtgatg ttcatataca gaagaatgcc
     5641 aacaatttac agatgatttt acaaattctt aattttcctt ttctaaatgg atttgttcac
     5701 acattatctc ttcaaagcaa ttgaattttc tggaaaagat tggctgtgtg tgtaaaaggt
     5761 gctactgagg aatttaatgt gacatggaat gaagcaaaca gcagcctaag acaggatggc
     5821 aggaaaaatg tcacatgttt aaaataagag tgaaaccaac gacctggact tatagagact
     5881 gtgtgtcctt aggcaaagct cttgtacaaa cagtgcttgt gactgtgaaa ttcggggttt
     5941 gcctggattg tgttaaatgt gtgcttggat gtatcataaa aaagaatcgg ttctctgtat
     6001 ttgacatttt ctgtaattta tctaaagtta caactggcta tgcctgattt ttatcactat
     6061 tgaatcaatc attttaaaga atgttaagtc tgctgatttg gagagctcat ttacatttat
     6121 tttgaatgtc tcttatttaa cacaactgat ttaagcttta gatctattac cctgttatcc
     6181 ctagggaaaa tagaatgaag tgatctccaa aaaataagta ctttttgact gtaaataaaa
     6241 ttgtaaggag taaaaagtac ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt
     6301 attgattttt aattgtactc aagtaaagta aaaatcccca aaaataatac ttaagtacag
     6361 taatcaagta aaattactca agtactttac acctctgggc ccaattcgcc c
//
