LOCUS       Exported                7194 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Myl7 promoter driving HA-tagged biotin ligase (BirA), a Thosea 
            asigna virus peptide (2A), and membrane-tethered mCherry protein 
            (membCherry); flanked by Tol2 sequences.
ACCESSION   .
VERSION     .
KEYWORDS    pMT-myl7-cytoBirA-2A-mCherry_Ras
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7194)
  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 7194)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7194
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..19
                     /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     8..30
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     27..43
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..43
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     62..82
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        64..82
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     119..135
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     119..135
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      2034..2043
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2157..3119
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3135..3188
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3195..3902
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3340..3358)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3608..3627
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(3988..4005)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4015..4033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4040..4174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4064..4083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4118..4137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(4444..4462)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4472..4489)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4472..4488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4481..4503)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4630..5085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4717..4736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4927..4948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5111..5215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5216..6076
                     /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(5434..5453)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6247..6835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6736..6755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6989..7006
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7123..7144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7159..7189
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
       61 cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgctc
      121 tagaactagt ggatctgctg ggcttgctga aggtaggggg tcaagaacca gaggtgtaaa
      181 gtacttgagt aattttactt gattactgta cttaagtatt atttttgggg atttttactt
      241 tacttgagta caattaaaaa tcaatacttt tacttttact taattacatt tttttagaaa
      301 aaaaagtact ttttactcct tacaatttta tttacagtca aaaagtactt attttttgga
      361 gatcacttgg gcccggctcg agtgtgacca aagcttaaat cagttgtgtt aaataagaga
      421 cattcaaaat aaatgtaaat gagctctcca aatcagcaga cttaacattc tttaaaatga
      481 ttgattcaat agtgatacaa atcaggcata gcagttgtaa cttttgataa attacagaaa
      541 atgtcaaata cggagaaccg attctttttt atgacacatc caagcacaca tttaacacaa
      601 tccaggcaaa ccccgaattt cagagtcaca agcactgttt gtacaagagc tttgcctaag
      661 gacgcacagt ctctatgagt ccaggtcgtt ggtttcactc ttattttaaa catgtgacat
      721 ttttcctgcc atcctgtctt aggctgctgt ttgcttcatt tcatgtcaca ttaatttcct
      781 cagtagcacc ttttacacac acagccaatc ttttccagaa cattcaattg ctttgaagag
      841 ataatgtgtg aacaaatcca tttagaaaag gaaaattaag aatttgtaaa atcatctgta
      901 aattgttggc attcttctgt atatgaacat cacatcattt acaggtaaag gtctggtcat
      961 taattatatg acaatttact ggtattattt tgtgaaaggg gctattttca atgcattcat
     1021 ccatcctttt catccctcaa atctctcatt cacgtccccc tccccatctg cacactttat
     1081 ctcattttcc accctgctgg aatctgagca cttgtgcagt tatcagggct cctgtattta
     1141 ggaggctctg ggtgtccatg taggggacga acagaaacac tgcagacctt tatagaagaa
     1201 caaatgataa gagtcctcat acataaagac tccattagaa acgtcagtga cccaggagcc
     1261 cagaccaaca gcaaagcaga cagtgaacgt ggtgagtaga caaagctata cttttttggt
     1321 tttgaatata atattaatgt gaaaataaaa agggtctata tgaagttaaa tggtgtttgt
     1381 ttgttgatat taaatattag aagcatcttt tctgcatttg tatgttgtga ttctatcttt
     1441 taatggtatt tttacttagg tacttttttc tgtacactgt aaaacgcaaa aaagttaagg
     1501 taactcaaat catttgagaa aaccgattgt aacaagccat ctgaagttca aaaactaatc
     1561 taaatgagta ctgtgaactt aatctatttg agtaaaagaa gcaatttgag cacagtaaaa
     1621 cccagtaaat gaagagaact caaccaactg agtactgtaa aacccaataa gttgaggcag
     1681 ctcacaccgt tagaggaaac cgattgctac aaaccatttg agttaaaaaa aagaatctat
     1741 atgagtactg tgaacttact ccatttaagt tgaagtaatg aggtaattaa ttaactcatt
     1801 accctcaaca ctaagttcaa aactcttttc aaatgagtag aattaatctt cagccaattt
     1861 tgagttaact atactcattt catttgataa agttgactat tgggttttac agtgtataaa
     1921 tgctataaat tgagtttcca aataatgctt ctcttttttt tttactcaca ggctagtcga
     1981 attcaaggtt aagcggccgc aagctagatt ggcattccgg tactgttggt aaagccacca
     2041 tggccaccta tgacgtccca gattacgcaa gtttgcccgg gtatgatgtt cctgattatg
     2101 ctagcctccc gggttacgat gtgcccgact atgcctcact tccaggcaga tctcctatga
     2161 aggataacac cgtgccactg aaattgattg ccctgttagc gaacggtgaa tttcactctg
     2221 gcgagcagtt gggtgaaacg ctgggaatga gccgggcggc tattaataaa cacattcaga
     2281 cactgcgtga ctggggcgtt gatgtcttta ccgttccggg taaaggatac agcctgcctg
     2341 agcctatcca gttacttaat gctaaacaga tattgggtca gctggatggc ggtagtgtag
     2401 ccgtgctgcc agtgattgac tccacgaatc agtaccttct tgatcgtatc ggagagctta
     2461 aatcgggcga tgcttgcatt gcagaatacc agcaggctgg ccgtggtcgc cggggtcgga
     2521 aatggttttc gccttttggc gcaaacttat atttgtcgat gttctggcgt ctggaacaag
     2581 gcccggcggc ggcgattggt ttaagtctgg ttatcggtat cgtgatggcg gaagtattac
     2641 gcaagctggg tgcagataaa gttcgtgtta aatggcctaa tgacctctat ctgcaggatc
     2701 gcaagctggc aggcattctg gtggagctga ctggcaaaac tggcgatgcg gcgcaaatag
     2761 tcattggagc cgggatcaac atggcaatgc gccgtgttga agagagtgtc gttaatcagg
     2821 ggtggatcac gctgcaggaa gcggggatca atctcgatcg taatacgttg gcggccatgc
     2881 taatacgtga attacgtgct gcgttggaac tcttcgaaca agaaggattg gcaccttatc
     2941 tgtcgcgctg ggaaaagctg gataatttta ttaatcgccc agtgaaactt atcattggtg
     3001 ataaagaaat atttggcatt tcacgcggaa tagacaaaca gggggcttta ttacttgagc
     3061 aggatggaat aataaaaccc tggatgggcg gtgaaatatc cctgcgtagt gcagaaaaag
     3121 atatcggatc aggagagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     3181 ccgggcccag atctatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     3241 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     3301 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3361 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     3421 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     3481 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     3541 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     3601 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     3661 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     3721 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     3781 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     3841 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     3901 agggaggagg aagatctaag ctgaaccctc ctgatgagag tggccccggc tgcatgagct
     3961 gcaagtgtgt gctctcctga tctagaacta tagtgagtcg tattacccgg actccctata
     4021 gtgagtcgta ttaaccggtg atccagacat gataagatac attgatgagt ttggacaaac
     4081 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4141 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4201 gtttcaggtt cagggggagg tgtgggaggt tttttgatat cccgggttta aacgcatgca
     4261 gatctaatac tcaagtacaa ttttaatgga gtactttttt acttttactc aagtaagatt
     4321 ctagccagat acttttactt ttaattgagt aaaattttcc ctaagtactt gtactttcac
     4381 ttgagtaaaa tttttgagta ctttttacac ctctgtcaag aaccatatgg gtacccaatt
     4441 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     4501 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4561 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4621 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4681 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4921 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4981 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5041 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     5101 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5161 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5221 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5281 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5341 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5401 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5461 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5521 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5581 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5641 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5701 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5761 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5821 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5881 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5941 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6001 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6061 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6121 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6181 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6241 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6301 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6361 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6421 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6481 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6541 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6601 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6661 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6721 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6781 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6841 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6901 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6961 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7021 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7081 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7141 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtgg
//
