LOCUS       62056_3395        7514 bp DNA     circular SYN 27-APR-2021
DEFINITION  Cloning vector pB-tetO2-mTET3cd-mCherry, complete sequence.
ACCESSION   MW139646
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Hore TA.
  TITLE     Pronounced specificity of the TET enzyme catalytic domain guides 
            cellular function
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7514)
  AUTHORS   Hore TA.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-OCT-2020) Department of Anatomy, University of Otago, 
            270 Great King Street, Dunedin, Otago 9016, New Zealand
REFERENCE   3  (bases 1 to 7514)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-OCT-2020) Department of Anatomy, University of Otago, 270 Great 
            King Street, Dunedin, Otago 9016, New Zealand"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Geneious v. March 2019
            Assembly Name         :: pB-tetO2-mTET3cd-mCherry
            Coverage              :: 463
            Sequencing Technology :: MiSeq
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   702..936
                     /label=LTR
                     /rpt_family="PiggyBAC LTR"
                     /note="LTR"
     protein_bind    1140..1158
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      join(1177..1195,1214..1232,1251..1269,1288..1306,1325..1343,
                     1362..1380,1399..1417)
                     /label=tetO2
                     /note="tetO2"
                     /regulatory_class="response_element"
     protein_bind    1543..1567
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1573..2922
                     /codon_start=1
                     /transl_table=11
                     /product="mTET3cd"
                     /label=mTET3cd
                     /protein_id="QQM12951.1"
                     /translation="MRPLEFPTCDCVEQIVEKDEGPYYTHLGSGPTVASIRELMEDRYG
                     EKGKAIRIEKVIYTGKEGKSSRGCPIAKWVIRRHTLEEKLLCLVRHRAGHHCQNAVIVI
                     LILAWEGIPRSLGDTLYQELTDTLRKYGNPTSRRCGLNDDRTCACQGKDPNTCGASFSF
                     GCSWSMYFNGCKYARSKTPRKFRLTGDNPKEEEVLRNSFQDLATEVAPLYKRLAPQAYQ
                     NQVTNEDVAIDCRLGLKEGRPFSGVTACMDFCAHAHKDQHNLYNGCTVVCTLTKEDNRC
                     VGQIPEDEQLHVLPLYKMASTDEFGSEENQNAKVSSGAIQVLTAFPREVRRLPEPAKSG
                     GGGSGGGGSGGGGSELWSDSEHNFLDENIGGVAVAPAHCSILIECARRELHATTPLKKP
                     NRCHPTRISLVFYQHKNLNQPNHGLALWEAKMKQLAERARQRQEEAARLG"
     protein_bind    complement(2924..2948)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2990..3576
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3577..4284
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     polyA_signal    4364..4419
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4867..5179
                     /label=LTR
                     /rpt_family="PiggyBAC LTR"
                     /note="LTR"
     promoter        complement(5324..5342)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5363..5379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5387..5403)
                     /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)."
     promoter        complement(5411..5441)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5456..5477)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5765..6353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6527..7384)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7385..7489)
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttaatta actcgaggga tccaagctgg ccgcacgtct
     1141 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     1201 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     1261 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     1321 gagctcccta tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga
     1381 agtcgacacg tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gtggcggcca tcacaagttt gtacaaaaaa
     1561 gcaggcttca ccatgcgccc tttagagttc cctacctgcg attgtgtcga acaaatagtg
     1621 gagaaagatg aaggcccata ttacactcac ctgggatctg gccccacagt agcttctatc
     1681 cgggaactca tggaggatcg gtatggagaa aaggggaaag ctatccggat tgagaaggtc
     1741 atctacacgg gcaaggaggg gaagagttct cgaggctgtc ccatcgccaa gtgggtgatc
     1801 cgaagacaca cactggagga gaagctgctg tgcctggtgc ggcatcgggc aggccaccat
     1861 tgtcagaacg ccgtgattgt tatcttgatc ctggcctggg agggcatccc tcgaagcctt
     1921 ggggacaccc tctaccagga gcttactgat accctccgga agtatggcaa ccctaccagc
     1981 cggagatgtg gcctcaatga tgaccggacc tgtgcttgcc aaggcaaaga ccctaacacc
     2041 tgcggtgcct ccttctcctt cggctgttcc tggagcatgt acttcaacgg ctgcaaatat
     2101 gctcggagca agacgccacg aaagttccgc ctcacgggag acaatccgaa ggaggaggag
     2161 gtgctccgga atagctttca ggatctggcc actgaagttg ctcccctcta caagcggctc
     2221 gcaccccagg cctatcagaa ccaggtgacc aatgaggatg tggcgatcga ctgccgcctg
     2281 gggctgaagg aagggagacc cttctcaggg gtcacagcct gcatggactt ctgtgcccac
     2341 gcccacaagg accaacataa cctctacaat gggtgcactg tggtctgcac cctgaccaag
     2401 gaagacaatc gctgcgtggg ccagatccct gaggacgagc aactgcacgt gctgcccctc
     2461 tacaagatgg ccagcacgga tgagtttggc agcgaggaaa accagaacgc caaggtcagt
     2521 agtggggcca tccaggtgct cacagcattc cccagagagg tccggcggct gcctgagcct
     2581 gccaagtccg gaggtggcgg gagtggcggc ggtgggtcgg gtggcggtgg cagcgaactg
     2641 tggtcggaca gtgaacacaa cttcctggat gagaacatag gcggggtggc cgtggccccc
     2701 gcccattgct ccatcctcat cgagtgtgcc cggcgagagc tgcatgccac cactccactc
     2761 aaaaaaccca accgctgcca ccccacccgc atctcgctgg tcttctacca acacaagaac
     2821 ctcaaccagc ccaaccacgg gctggcgctc tgggaggcca agatgaagca gctggcggaa
     2881 cgggcgcggc agcggcaaga ggaggccgca cgcctgggct aggacccagc tttcttgtac
     2941 aaagtggtga tctagctaaa acgcggcctc gaatcgaatt ccgccccccc ccctctccct
     3001 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     3061 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     3121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     3181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     3241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     3301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     3361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     3421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     3481 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     3541 tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggaggat
     3601 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     3661 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3721 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3781 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3841 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3901 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3961 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     4021 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     4081 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     4141 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     4201 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     4261 ggcggcatgg acgagctgta caagtaaagc ggccgcgatc tttttccctc tgccaaaaat
     4321 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4381 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     4441 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     4501 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     4561 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     4621 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     4681 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     4741 agatccctcg acctgcagga attcgatatc aagctctagc caacaagctc gtcatcgctt
     4801 tgcagaagag cagagaggat atgctcatcg tctaaagaac tacccatttt attatatatt
     4861 agtcacgata tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat
     4921 gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat
     4981 gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac
     5041 aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga
     5101 ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag
     5161 actatctttc tagggttaaa aaagatttgc gctttactcg acctaaactt taaacacgtc
     5221 atagaatctt cgtttgacaa aaaccacatt gtggccaagc tgtgtgacgc gacgcgcgct
     5281 aaagaatggc aaaccaagtc gcgcgaggta ccagcttttg ttccctttag tgagggttaa
     5341 tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc acct
//
