LOCUS       sequence-149739-        7692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Beta-actin2 promoter driving N-term Avi-tagged protein containing 
            Cerulean protein fused to the carboxy-terminal domain of avian Ran 
            GTPase-activating protein 1 (RanGap1); flanked by Tol2 sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7692)
  AUTHORS   Trinh LA, Chong-Morrison V, Gavriouchkina D, Hochgreb-Hagele T, 
            Senanayake U, Fraser SE, Sauka-Spengler T
  TITLE     Biotagging of Specific Cell Populations in Zebrafish Reveals Gene 
            Regulatory Logic Encoded in the Nuclear Transcriptome.
  JOURNAL   Cell Rep. 2017 Apr 11;19(2):425-440. doi: 
            10.1016/j.celrep.2017.03.045.
  PUBMED    28402863
REFERENCE   2  (bases 1 to 7692)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-04-11- 11"; volume: "19"; issue: "2"; pages: "425-440"
COMMENT     Sequence Label: pMTB2-Avi-Cerulean-RanGAP
FEATURES             Location/Qualifiers
     source          1..7692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      353..941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     842..861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1095..1112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1229..1250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1265..1270,1271..1288,1289..1295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1319
                     /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     1308..1330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1327..1343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1327..1343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1362..1382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1364..1382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1419..1435
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1419..1435
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(1469..1480,1481..1668)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        4383..4401
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4383..4400
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      4436..4445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4451..4495
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4502..4522
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4526..4528,4529..4531,4532..5242)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4571..4592)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4832..4851)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5179..5200
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5811..5830)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5812..5830)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5837..5971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5861..5880)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5915..5934
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(6063..6200,6201..6212)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6270..6287)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6270..6286)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6279..6301)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6428..6883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6515..6534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6725..6746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6909..7013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7014..7082,7083..7692,1..182)
                     /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(7232..7251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
       61 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      481 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1021 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1081 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     1141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1201 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1261 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1321 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     1381 gggaacaaaa gctggagctc caccgcggtg gcggccgctc tagaactagt ggatctgctg
     1441 ggcttgctga aggtaggggg tcaagaacca gaggtgtaaa gtacttgagt aattttactt
     1501 gattactgta cttaagtatt atttttgggg atttttactt tacttgagta caattaaaaa
     1561 tcaatacttt tacttttact taattacatt tttttagaaa aaaaagtact ttttactcct
     1621 tacaatttta tttacagtca aaaagtactt attttttgga gatcacttgg gcccggctcg
     1681 agggcctagt cattacctat actcaacgtt tactctgaga aacttccata ttttagaggt
     1741 aaaatgctca gaattgtaaa aagtcttgtc aaaatacttg gaatttgcca aaaacaaaat
     1801 aaaagtcaca tcacctcatt gtccttggta tcagcaagca cttttaacac aatcagctgt
     1861 ttttgtttat aactcccact gaaggccatg tgcattactc ccaagttaca cagtaaaaag
     1921 tcacatgtag tttatcatga tcactgctgc tcaagcacag ctttatctgt tattcagtgt
     1981 ggcaatatct gaaatgtgtg caaaatatta tttattgtag ctgatcacca acactttcag
     2041 tgtttttttt ttctactttt aatgagaatt atgttcttag gatttatcct tataaacctc
     2101 tttaaaaaga aatgttatgt ctgcatagac acgacatgaa attatgaaca gaaaatttaa
     2161 ctatttaaga tcaagacttg acatataaca tttctaatga acattaaacc tgaggtaggc
     2221 ccacttgcaa aaatgtaact gctatcactc actttataca tcattgtgta acttgtacac
     2281 acaaaaatag tgaagcaatg tggcatggaa tgcaggtccc attcattcaa agttattaaa
     2341 gcactttctt aaagtgcatt actactgagc taagaccagc tagttgctca tacaacatac
     2401 tacaagtagt gtcatcctaa tttggataaa aaaaatgtcc attttataca tggaaaagta
     2461 ctcattagat cccccattgt ttgtattacg gtatttcgtg aacacaagag gtaaatgacc
     2521 tacagagctg ctgttgtgtt agattgaaaa cacaacacag gatcatggag gcatccatca
     2581 ttatgacgca ttctgccact tgtaactctg cacagacttt ttgaaaagtt taattgagtc
     2641 ttggcgttgt cgcagaaaaa tacgaaaaat tctgtgttaa aatttgcaaa taaaaattcc
     2701 agttttatag gaaatcgcca tggccttgtt ctttaaccag ttaagcttcc ccttctttca
     2761 ctctcaagtt gcaagaagca agtgtagcaa tgtgcacgcg acagccgggt gtgtgacgct
     2821 ggaccaatca gagcacagag ctccgaaagt ttacctttta tggctagagc cgggtatgtg
     2881 ccgtcatata aaagagcgcg cccagctttt cagcctcact ttgagctcct ccacacgcag
     2941 ctagtgcgga atatcatctg cttgtaaccc attctcttaa gtcgacaacc ccccaaaccc
     3001 aaggtgagtt ggttttacag tttttaagcg tttaattata gtttctatct attaatgtta
     3061 agtaaaatgt gtaaatggat aaatatgatt tgatgaagtc tttagacagt tcataacggt
     3121 ctctttgtag ttatctacta ggtttgattt ataagtggtg gttcttttga taatctatgt
     3181 tacatttaat aaatattggc tgtatcttaa cttttttgta gcgttagtgg atgtggcagg
     3241 tgagaatgtc ggtaatttaa cgtgaccagt cgtaggcacg acattgaaca ttgaataggc
     3301 cggtgtgaaa tgggtgttaa gtcttaatat aaagttgtgt tctggctaat ctctctttaa
     3361 cagccataaa tgtgattttt aactacagtt ttgagggtgc tggtgagcat gttgcacact
     3421 tgatggatag ccggcatggg aagttctttg tgcaggcagt gctgcagcag ggtgtgacct
     3481 actttagcta gccggctaac cagctctcat ctgatgtaac gtaaacccca tcaatctaac
     3541 atcggctcat tcgtgcttta taagatcgaa gatcttgtta ttagtaacgg gtattggcat
     3601 ttagttttaa cttggttaat tttctttttg ttatgtgttc tatttaaatg tagcctgcag
     3661 ccccctctgt ctttaatgaa gctgaagatt aacgttaacg ttagtgaagt gttgtatccg
     3721 ctatgaaaga tttgaatttt agaggtgtgg tccactttaa taacggctgc ggagttctct
     3781 ggctgcactt cagccgctgt gactgtcgaa gcagagccca tcagtttttc aggatctcgg
     3841 tgatttttga ggctagacta tgaactgaac cgactgagag tgaccgtcca gaaatgcttt
     3901 agtgtgaagt ctccgctcac gaggagtccg tggactgact gcagatctgt gacgcagtaa
     3961 cacccggcgg cagacacccg ttgaaatgcg gttgtgtaat tgatcgcagg cgagggtgga
     4021 agaggatgtg aaacttcatt tgtgtaaaat ttagggagtg gtcctggtgt gatgaatgtc
     4081 gaaatctgtt cctttttact gagccctacg actctggctg agtgccacac cgccggcagc
     4141 cgcaaagcgg ctcaaaccat tgccttttat ggtaataatg agagaatgca gagggacttc
     4201 ctttgtctgg catatctgag gcgcgcgctg tcactagcgc ccaccagcgg tcagactgta
     4261 gaatgcagag caaaacagga agttgactcc agatggtcac atgcttgctg aaacgcctct
     4321 ccctggcagc agtgcaattc taacagtttc cctttctttt acagttcagc catcctaggc
     4381 ctatttaggt gacactatag aagagtacaa gctacttgtt ctttttgcag gatccgccac
     4441 catggctggt ggcctgaatg acatctttga ggcccagaag atcgagtggc atgagggagg
     4501 agagaacctg tacttccagg gaggaatggt gagcaagggc gaggagctgt tcaccggggt
     4561 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4621 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     4681 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt
     4741 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4801 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4861 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4921 ggaggacggc aacatcctgg ggcacaagct ggagtacaac gccatcagcg acaacgtcta
     4981 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     5041 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     5101 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     5161 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     5221 cggcatggac gagctgtaca aggccgctgc atcgattatg gctcctaaga agataattgg
     5281 ttcacaggat tcaactccag tgccatctcc tcctgtggac gttgccacgt tccttgcttt
     5341 cccatcacca gagaagctgc tgcggctagg gccaaagtgt tctgtgctga tagctcagca
     5401 gacagataca tctgatgtag agaaagtagt tacagctctc ttaaggatat cttcagtctt
     5461 caaagatgaa gcccaagtaa aaacagcagt gcatgaaaca gcagatgcct tgatgaaaaa
     5521 agccttctct tccgccacat ttaattcaga tgcattcatc acaagactct tgatccacat
     5581 ggggctactt aagagtgaag agaagatcaa agctgtcccg agcctctatg gtattcttat
     5641 gactttgaat catatggtcc agcaggatta tttccctaaa tcactggccc cagttctctc
     5701 tgcttttgtc acaaaaccaa accgtgctct cgactcctgt tcctttgccc gccacatgct
     5761 tttacaaacc ctccaccagc tgtaataatg aattcatacg tatccggact ccctatagtg
     5821 agtcgtatta accggtgatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     5881 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     5941 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     6001 tcaggttcag ggggaggtgt gggaggtttt ttgatatccc gggtttaaac gcatgcagat
     6061 ctaatactca agtacaattt taatggagta cttttttact tttactcaag taagattcta
     6121 gccagatact tttactttta attgagtaaa attttcccta agtacttgta ctttcacttg
     6181 agtaaaattt ttgagtactt tttacacctc tgtcaagaac catatgccgg tacccaatcg
     6241 ccctatagga cgtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     6301 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6361 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6421 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6481 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6541 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6601 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6661 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6721 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6781 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6841 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     6901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7081 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7141 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7201 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7261 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7321 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7381 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7441 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7501 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7561 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7621 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7681 ttccggctgg ct
//
