LOCUS       sequence-149785-        8566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitin promoter driving Avi-tagged protein containing Cerulean 
            protein fused to zebrafish Rpl10a ribosomal unit (Tryon et al. 
            2012); flanked by Tol2 sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8566)
  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 8566)
  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-Rpl10a
FEATURES             Location/Qualifiers
     source          1..8566
                     /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)"
     CDS             5174..5218
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5225..5245
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(5249..5251,5252..5254,5255..5965)
                     /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(5294..5315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5555..5574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5902..5923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             6049..6057
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(6685..6704)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6686..6704)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6711..6845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6735..6754)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6789..6808
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(6937..7074,7075..7086)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7144..7161)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7144..7160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7153..7175)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7302..7757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7389..7408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7599..7620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7783..7887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7888..7956,7957..8566,1..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(8106..8125)
                     /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 aaccagcaaa gttctagaat ttgtcgaaac atttatgtta tatatttcct gaaaaaaatt
     1741 ctgagtaagt tcttaagtgt tattgccagc aacataaaca acagacggca aaatgaataa
     1801 atgataacaa agcagtaggc ttaaataaac ctaattttta taggctgttc tctacaaccc
     1861 tcaaacagtg attagttttg tacttataaa cttgcccttt cattcatatt tcaagaaaat
     1921 tggttcagaa gatctggata ttctagcagt tgttcaagct catggaggga tcagtgacct
     1981 gattccacaa tgactaggcc taatccagaa attagatgac tgtcaacata aaaaggcaca
     2041 gcactcacta gctgccctat atattttatt atattttaca tatattattt tatttattta
     2101 gctctgagtg ctgtactttc tggttaaaga aaactgctta caacagctaa cctgtactac
     2161 ctcaggctca gggaatttgg aacaggtttg tctggtttgt ttctttaacc atgcatgctt
     2221 gttttcaact atggcaacac agtcacatgg gacattacag aaatgatttg tcgatgacat
     2281 gcgacttttc tttaataaag cgcaaagatc ccaaaaagca aacttttaac aaaaatcata
     2341 taattatatt ttcaatccag ctttgtagca actttgtgct gctgttcact cagcaacaga
     2401 tagtcagtat aaggtcagtg tgtctcaaag cagtgccatc tgtttcacac attgcgttct
     2461 atatataagt gtgctggttg acacgacact gtataaggcc taggctaaaa cacaaacaat
     2521 gtagaatgac actgtgtttt ttttgtaaac aaatgttgtt tttggttaaa catctttgtg
     2581 aaaacatcct cctgtcatgt atttgctata ttcaaatgtt aaacccgtgc agaatagaac
     2641 atatacaaaa aaaaacaaca caacacattt ttaaacatta ttaaatatca agtattgctg
     2701 gcagttctgt ttctgtttta cagtaccctt tgccacagtt ctccgctttt cctggtccag
     2761 attccacaag tctgattcac caatagcaaa gcgaataaac aaccaaagca gccaatcact
     2821 gcttgtagac tgtcctgcga gaccggccca ttccagcaca ttctggaaac ttcctttata
     2881 tgataattat aaatacattt aaattattga tacaaaacat gtaattccta gaacataacc
     2941 atagcaatca ttagttttca gggtaattat gtatttttag gatttgactg cggaaagatc
     3001 tggtcatgtg acgtctcatg aacgtcacgg ccctgggttt ctataaatac agtaggactc
     3061 tcgaccatcg gcagattttt cgaagaagaa gatcagtttc aggagccgta ctgttccgtt
     3121 tcaacgcaaa tattaacggt aagagcgaat ttcctagttt gttttcatgc cattctttaa
     3181 aaccatagcg tattacttta attatagtaa actttcgctt tctttattac aagagacgtt
     3241 ttgtgttgat tctccgcgga cattttcggt cagacaatca gaaaatgacc gcggaggacc
     3301 agtaacttgc attacacgta agttaaatct tcgtgtatta aaatggttag gttgttaacg
     3361 tcaaataggt taccgtgttt gcgtgtgatc aggttggttt tgttagattt ttgtcagtat
     3421 ttttaattta tttgttttag tttatttatt ttttttgctg aatcatagtt tgtgaacaaa
     3481 gaacccggat gttacataca gtacagccgc catgttacag agagttataa cttaatcatt
     3541 ttaaaaataa ttttgcctta cttttagttt gtcatgttga gaaatgagga aatgttaaaa
     3601 tgaggaaata tccaattaat ttaatatatc aaaataatcc atgattacaa tgcactgaac
     3661 tggagaaaat taagatgttt tctagtgtca tgaaacaaat gtaagagatg tacattgtag
     3721 atgttttatg tcaagaattg gctagttgat gcagcatact ggcgatactc agttgtaata
     3781 acagtaacgt tacatgttaa tagactactg agtatgctgt tctgtctatg tatgctctgt
     3841 aagctacgag aaggactttt ttaaacagta aagggtgcaa tatttttaca aattgaatta
     3901 aataaaggct gtctattaag taatatgctt gatatttttc ttacttgatc ggaaataaga
     3961 aaaaatataa acgttgttgc tctaaaaatc ctagttcagt ttagccaacc acaaatacct
     4021 ttttgttcct ccaacagttt ttttttcttc tctataatat ttggcagtct atagtactcc
     4081 aaatgtttcc ccacagtcta actaattggt acagccaaaa tcatgacact tattgcaata
     4141 ataattttgg ttcattggca ttgttgatag cctgtgccac taatatggtc gattgatcat
     4201 gcttcaggaa gaaaactata ttgtttgatg taagattatt aaatcttcac ctgcttccat
     4261 tacaaactat tcccatctta ttgaattctg gtatgtctta aaggattagt tcacttccca
     4321 aatcaaaatt tacttagcct tttttcatcc atgatccctt tttttcatca ttaatgaaga
     4381 aattgttttt gaaaaagttt caagattttt ttctctatat tgtggagctt gttagtttaa
     4441 aattccaaaa tgcaatatgt ggcttcaaat ggttctaaat gatcccagtc aaggaataac
     4501 agtcttatct aatgaaacca ttagaccttt aaaaataaaa aaaataaaag tatttatttt
     4561 taaatgactg agtgattaag ttgaatttca gcgtttcctt actgtgtaga agtccttcct
     4621 tactggcccc accctttggt tctctgccaa tctgctacct aatgtaatgt tgtggaacat
     4681 tattattctt tatttcttaa ttttttattt tttattttaa aacaatgtaa actgcacaga
     4741 tgtgcagttt gtttaaaatg gccaatgctt tggaaatgca tgacataatt agatttcatg
     4801 atgcacaaag ccaaatctca gagcttgtgc aaaatgagct atcatttcac taggtaagac
     4861 cctaaatttt catataggat catttggaca attttgctgc aggtaaaatg cattctatag
     4921 tccactgtca gccattgttt tggatagtat ttatttttct ctacaagtat agtcaatagt
     4981 tttctattat tttaaaggtt tgtaacattt aagggtgacc aaatgcaaag taaaatttca
     5041 ttttcgggtg aactatctcg tttaacatgg gagaagtgca aaacatacat tattggctag
     5101 aacattgtag tattttttaa atggaaatgt gtgattgcta atcttacttt gaatttgttt
     5161 acagatggct ggtggcctga atgacatctt tgaggcccag aagatcgagt ggcatgaggg
     5221 aggagagaac ctgtacttcc agggaggaat ggtgagcaag ggcgaggagc tgttcaccgg
     5281 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     5341 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     5401 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg
     5461 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     5521 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     5581 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     5641 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacgccatca gcgacaacgt
     5701 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     5761 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     5821 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     5881 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     5941 tctcggcatg gacgagctgt acaaggccgc tgcatcgggc cggactcaga tctcgagctc
     6001 aagcttcgaa ttcagcaagg tctcgagaga cacgttgtac gaggccgtta aggaggtcca
     6061 ggccggttcc cgtcgcaaga agagaaagtt tcttgaaacc gtggaactcc agatcagctt
     6121 gaagaactac gaccctcaga aggacaagcg tttctcaggc actgtcaggc tgaagaccac
     6181 cccgcggccc aagttctctg tctgcgttct tggagaccag cagcattgtg atgaagcaaa
     6241 ggccgctgaa ctgccacaca tggacattga ggctcttaag aagctcaata agaacaagaa
     6301 actggtcaag aagctggcaa agaagtacga tgctttcctg gcctctgagt ctctgatcaa
     6361 gcagattcct cgtatcctgg gccctggcct caacaaggca ggaaagtttc cctctctgct
     6421 cacccataat gagaacctgg gcaccaaggt ggatgaggtg aaatccacca tcaaattcca
     6481 gatgaagaag gttctgtgtc tggctgtggc cgtgggtcat gtgaagatga gtgaggaaga
     6541 gctggtctac aacatccacc tggctgtcaa cttcctggtg tctctgctga agaagaactg
     6601 gcaaaacgtc agagctcttt acatcaagag caccatggga aaaccacagc gcctctacta
     6661 gtagaattca tacgtatccg gactccctat agtgagtcgt attaaccggt gatccagaca
     6721 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     6781 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     6841 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     6901 ttttttgata tcccgggttt aaacgcatgc agatctaata ctcaagtaca attttaatgg
     6961 agtacttttt tacttttact caagtaagat tctagccaga tacttttact tttaattgag
     7021 taaaattttc cctaagtact tgtactttca cttgagtaaa atttttgagt actttttaca
     7081 cctctgtcaa gaaccatatg ccggtaccca atcgccctat aggacgtcgt attacgcgcg
     7141 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     7201 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     7261 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     7321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     7381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     7441 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     7501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     7681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     7741 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     7801 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     7861 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     7921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     7981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     8041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     8101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     8161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     8221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     8281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     8341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     8401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     8461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     8521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggct
//
