LOCUS       sequence-149777-        8428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitin promoter driving 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 8428)
  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 8428)
  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: pMT-ubb-Avi-Cerulean-RanGAP
FEATURES             Location/Qualifiers
     source          1..8428
                     /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)"
     regulatory      5172..5181
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5187..5231
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5238..5258
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(5262..5264,5265..5267,5268..5978)
                     /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(5307..5328)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5568..5587)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5915..5936
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6547..6566)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6548..6566)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6573..6707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6597..6616)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6651..6670
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(6799..6936,6937..6948)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7006..7023)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7006..7022)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7015..7037)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7164..7619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7251..7270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7461..7482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7645..7749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7750..7818,7819..8428,1..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(7968..7987)
                     /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 agaccagcaa agttctagaa tttgtcgaaa catttatgtt atatatttcc tgaaaaaaat
     1741 tctgagtaag ttcttaagtg ttattgccag caacataaac aacagacggc aaaatgaata
     1801 aatgataaca aagcagtagg cttaaataaa cctaattttt ataggctgtt ctctacaacc
     1861 ctcaaacagt gattagtttt gtacttataa acttgccctt tcattcatat ttcaagaaaa
     1921 ttggttcaga agatctggat attctagcag ttgttcaagc tcatggaggg atcagtgacc
     1981 tgattccaca atgactaggc ctaatccaga aattagatga ctgtcaacat aaaaaggcac
     2041 agcactcact agctgcccta tatattttat tatattttac atatattatt ttatttattt
     2101 agctctgagt gctgtacttt ctggttaaag aaaactgctt acaacagcta acctgtacta
     2161 cctcaggctc agggaatttg gaacaggttt gtctggtttg tttctttaac catgcatgct
     2221 tgttttcaac tatggcaaca cagtcacatg ggacattaca gaaatgattt gtcgatgaca
     2281 tgcgactttt ctttaataaa gcgcaaagat cccaaaaagc aaacttttaa caaaaatcat
     2341 ataattatat tttcaatcca gctttgtagc aactttgtgc tgctgttcac tcagcaacag
     2401 atagtcagta taaggtcagt gtgtctcaaa gcagtgccat ctgtttcaca cattgcgttc
     2461 tatatataag tgtgctggtt gacacgacac tgtataaggc ctaggctaaa acacaaacaa
     2521 tgtagaatga cactgtgttt tttttgtaaa caaatgttgt ttttggttaa acatctttgt
     2581 gaaaacatcc tcctgtcatg tatttgctat attcaaatgt taaacccgtg cagaatagaa
     2641 catatacaaa aaaaaacaac acaacacatt tttaaacatt attaaatatc aagtattgct
     2701 ggcagttctg tttctgtttt acagtaccct ttgccacagt tctccgcttt tcctggtcca
     2761 gattccacaa gtctgattca ccaatagcaa agcgaataaa caaccaaagc agccaatcac
     2821 tgcttgtaga ctgtcctgcg agaccggccc attccagcac attctggaaa cttcctttat
     2881 atgataatta taaatacatt taaattattg atacaaaaca tgtaattcct agaacataac
     2941 catagcaatc attagttttc agggtaatta tgtattttta ggatttgact gcggaaagat
     3001 ctggtcatgt gacgtctcat gaacgtcacg gccctgggtt tctataaata cagtaggact
     3061 ctcgaccatc ggcagatttt tcgaagaaga agatcagttt caggagccgt actgttccgt
     3121 ttcaacgcaa atattaacgg taagagcgaa tttcctagtt tgttttcatg ccattcttta
     3181 aaaccatagc gtattacttt aattatagta aactttcgct ttctttatta caagagacgt
     3241 tttgtgttga ttctccgcgg acattttcgg tcagacaatc agaaaatgac cgcggaggac
     3301 cagtaacttg cattacacgt aagttaaatc ttcgtgtatt aaaatggtta ggttgttaac
     3361 gtcaaatagg ttaccgtgtt tgcgtgtgat caggttggtt ttgttagatt tttgtcagta
     3421 tttttaattt atttgtttta gtttatttat tttttttgct gaatcatagt ttgtgaacaa
     3481 agaacccgga tgttacatac agtacagccg ccatgttaca gagagttata acttaatcat
     3541 tttaaaaata attttgcctt acttttagtt tgtcatgttg agaaatgagg aaatgttaaa
     3601 atgaggaaat atccaattaa tttaatatat caaaataatc catgattaca atgcactgaa
     3661 ctggagaaaa ttaagatgtt ttctagtgtc atgaaacaaa tgtaagagat gtacattgta
     3721 gatgttttat gtcaagaatt ggctagttga tgcagcatac tggcgatact cagttgtaat
     3781 aacagtaacg ttacatgtta atagactact gagtatgctg ttctgtctat gtatgctctg
     3841 taagctacga gaaggacttt tttaaacagt aaagggtgca atatttttac aaattgaatt
     3901 aaataaaggc tgtctattaa gtaatatgct tgatattttt cttacttgat cggaaataag
     3961 aaaaaatata aacgttgttg ctctaaaaat cctagttcag tttagccaac cacaaatacc
     4021 tttttgttcc tccaacagtt tttttttctt ctctataata tttggcagtc tatagtactc
     4081 caaatgtttc cccacagtct aactaattgg tacagccaaa atcatgacac ttattgcaat
     4141 aataattttg gttcattggc attgttgata gcctgtgcca ctaatatggt cgattgatca
     4201 tgcttcagga agaaaactat attgtttgat gtaagattat taaatcttca cctgcttcca
     4261 ttacaaacta ttcccatctt attgaattct ggtatgtctt aaaggattag ttcacttccc
     4321 aaatcaaaat ttacttagcc ttttttcatc catgatccct ttttttcatc attaatgaag
     4381 aaattgtttt tgaaaaagtt tcaagatttt tttctctata ttgtggagct tgttagttta
     4441 aaattccaaa atgcaatatg tggcttcaaa tggttctaaa tgatcccagt caaggaataa
     4501 cagtcttatc taatgaaacc attagacctt taaaaataaa aaaaataaaa gtatttattt
     4561 ttaaatgact gagtgattaa gttgaatttc agcgtttcct tactgtgtag aagtccttcc
     4621 ttactggccc caccctttgg ttctctgcca atctgctacc taatgtaatg ttgtggaaca
     4681 ttattattct ttatttctta attttttatt ttttatttta aaacaatgta aactgcacag
     4741 atgtgcagtt tgtttaaaat ggccaatgct ttggaaatgc atgacataat tagatttcat
     4801 gatgcacaaa gccaaatctc agagcttgtg caaaatgagc tatcatttca ctaggtaaga
     4861 ccctaaattt tcatatagga tcatttggac aattttgctg caggtaaaat gcattctata
     4921 gtccactgtc agccattgtt ttggatagta tttatttttc tctacaagta tagtcaatag
     4981 ttttctatta ttttaaaggt ttgtaacatt taagggtgac caaatgcaaa gtaaaatttc
     5041 attttcgggt gaactatctc gtttaacatg ggagaagtgc aaaacataca ttattggcta
     5101 gaacattgta gtatttttta aatggaaatg tgtgattgct aatcttactt tgaatttgtt
     5161 tacagggatc cgccaccatg gctggtggcc tgaatgacat ctttgaggcc cagaagatcg
     5221 agtggcatga gggaggagag aacctgtact tccagggagg aatggtgagc aagggcgagg
     5281 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     5341 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     5401 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     5461 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     5521 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     5581 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     5641 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacgcca
     5701 tcagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     5761 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     5821 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     5881 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     5941 ccgccgggat cactctcggc atggacgagc tgtacaaggc cgctgcatcg attatggctc
     6001 ctaagaagat aattggttca caggattcaa ctccagtgcc atctcctcct gtggacgttg
     6061 ccacgttcct tgctttccca tcaccagaga agctgctgcg gctagggcca aagtgttctg
     6121 tgctgatagc tcagcagaca gatacatctg atgtagagaa agtagttaca gctctcttaa
     6181 ggatatcttc agtcttcaaa gatgaagccc aagtaaaaac agcagtgcat gaaacagcag
     6241 atgccttgat gaaaaaagcc ttctcttccg ccacatttaa ttcagatgca ttcatcacaa
     6301 gactcttgat ccacatgggg ctacttaaga gtgaagagaa gatcaaagct gtcccgagcc
     6361 tctatggtat tcttatgact ttgaatcata tggtccagca ggattatttc cctaaatcac
     6421 tggccccagt tctctctgct tttgtcacaa aaccaaaccg tgctctcgac tcctgttcct
     6481 ttgcccgcca catgctttta caaaccctcc accagctgta ataatgaatt catacgtatc
     6541 cggactccct atagtgagtc gtattaaccg gtgatccaga catgataaga tacattgatg
     6601 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     6661 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     6721 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttga tatcccgggt
     6781 ttaaacgcat gcagatctaa tactcaagta caattttaat ggagtacttt tttactttta
     6841 ctcaagtaag attctagcca gatactttta cttttaattg agtaaaattt tccctaagta
     6901 cttgtacttt cacttgagta aaatttttga gtacttttta cacctctgtc aagaaccata
     6961 tgccggtacc caatcgccct ataggacgtc gtattacgcg cgctcactgg ccgtcgtttt
     7021 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     7081 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     7141 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7201 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7261 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7321 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     7381 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7441 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7501 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     7561 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     7621 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7681 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7741 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7801 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7861 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7921 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7981 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     8041 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     8101 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     8161 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     8221 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     8281 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     8341 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8401 cttctgcgct cggcccttcc ggctggct
//
