LOCUS       7xTRE-tDTomato          5533 bp DNA     circular SYN 07-SEP-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5533)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 7, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..5533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      927..1515
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
                     /note="color: #ffff00"
     CDS             1698..1754
                     /codon_start=1
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=5.3"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="APNTQTASPRALADSLMQL"
     repeat_region   1748..1881
                     /label=AAV2 ITR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=95.0"
                     /note="/other=repeat_region"
                     /note="color: #a6acb3"
     repeat_region   1752..1881
                     /label=AAV2 ITR
                     /note="color: #ffcc99"
     CDS             1952..2020
                     /codon_start=1
                     /label=large T antigen (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=98.6"
                     /note="/match_length=3.2"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="YHICRGFTCFKKPPTSPPEPET"
     polyA_signal    2043..2164
                     /label=SV40 poly(A) signal
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=polyA_signal"
                     /note="color: #a6acb3"
     misc_feature    complement(2193..2212)
                     /label=MCS (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=18.5"
                     /note="/other=misc_feature"
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     2194..2210
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    complement(2213..2801)
                     /label=WPRE
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=misc_feature"
                     /note="color: #a6acb3; direction: LEFT"
     CDS             2319..2330
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 2330"
                     /translation="IEGR"
     CDS             complement(2815..4245)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             complement(2818..4245)
                     /codon_start=1
                     /label=tdTomato
                     /note="pLannotate"
                     /note="/database=fpbase"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     regulatory      4242..4251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     promoter        complement(4262..4574)
                     /label=tight TRE promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=99.4"
                     /note="/other=promoter"
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(4286..4324)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    4336..4354
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4372..4390
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4407..4425
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4443..4461
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4479..4497
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4514..4532
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    4550..4568
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     repeat_region   4642..4774
                     /label=AAV2 ITR (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=94.3"
                     /note="/other=repeat_region"
                     /note="color: #a6acb3"
     repeat_region   4642..4771
                     /label=AAV2 ITR
                     /note="color: #ffcc99"
     CDS             4769..4838
                     /codon_start=1
                     /label=lacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=40.2"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="QLA**RRGPHRSPFPTVAQPEWR"
     rep_origin      4843..5298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     rep_origin      4843..5271
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
                     /note="color: #ffff00"
     promoter        5324..5428
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(5429..5533,1..756)
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /note="color: #ccffcc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
       61 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      121 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      181 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      241 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      301 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      361 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      421 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
      481 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      541 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      601 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      661 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      721 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
      781 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
      841 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
      901 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
      961 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1021 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1081 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1141 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1201 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1261 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1321 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1381 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1441 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1501 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1561 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1621 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1681 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     1741 cattaatgca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     1801 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     1861 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
     1921 agccatgctc taggaagatc cttatcgatt ttaccacatt tgtagaggtt ttacttgctt
     1981 taaaaaacct cccacatctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2041 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2101 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2161 cttatcatgt ctgctcgaag cggccgcccg ggtcgaggtc gacggtatcg atgcggggag
     2221 gcggcccaaa gggagatccg actcgtctga gggcgaaggc gaagacgcgg aagaggccgc
     2281 agagccggca gcaggccgcg ggaaggaagg tccgctggat tgagggccga agggacgtag
     2341 cagaaggacg tcccgcgcag aatccaggtg gcaacacagg cgagcagcca aggaaaggac
     2401 gatgatttcc ccgacaacac cacggaattg tcagtgccca acagccgagc ccctgtccag
     2461 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa tagggagggg gaaagcgaaa
     2521 gtcccggaaa ggagctgaca ggtggtggca atgccccaac cagtgggggt tgcgtcagca
     2581 aacacagtgc acaccacgcc acgttgcctg acaacgggcc acaactcctc ataaagagac
     2641 agcaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga
     2701 tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata
     2761 ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatcgataag cttcttactt
     2821 gtacagctcg tccatgccgt acaggaacag gtggtggcgg ccctcggagc gctcgtactg
     2881 ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc agcttggtgt ccacgtagta
     2941 gtagccgggc agttgcacgg gcttcttggc catgtagatg gtcttgaact ccaccaggta
     3001 gtggccgccg tccttcagct tcagggcctg gtggatctcg cccttcagca cgccgtcgcg
     3061 ggggtacagg cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc
     3121 gtcggggggg aagttggtgc cgcgcatctt caccttgtag atcagcgtgc cgtcctgcag
     3181 ggaggagtcc tgggtcacgg tcaccagacc gccgtcctcg aagttcatca cgcgctccca
     3241 cttgaagccc tcggggaagg acagcttctt gtaatcgggg atgtcggcgg ggtgcttcac
     3301 gtacgccttg gagccgtaca tgaactgggg ggacaggatg tcccaggcga agggcagggg
     3361 gccgcccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc
     3421 ctcgccctcg atctcgaact cgtggccgtt catggagccc tccatgcgca ccttgaagcg
     3481 catgaactct ttgatgacgg ccatgttgtt gtcctcggag gaggcggtgc cggagctgcc
     3541 gctgccggtg ctgccggtgc catgccccag gaacaggtgg tggcggccct cggagcgctc
     3601 gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tggtgtccac
     3661 gtagtagtag ccgggcagtt gcacgggctt cttggccatg tagatggtct tgaactccac
     3721 caggtagtgg ccgccgtcct tcagcttcag ggcctggtgg atctcgccct tcagcacgcc
     3781 gtcgcggggg tacaggcgct cggtggaggc ctcccagccc atggtcttct tctgcattac
     3841 ggggccgtcg ggggggaagt tggtgccgcg catcttcacc ttgtagatca gcgtgccgtc
     3901 ctgcagggag gagtcctggg tcacggtcac cagaccgccg tcctcgaagt tcatcacgcg
     3961 ctcccacttg aagccctcgg ggaaggacag cttcttgtaa tcggggatgt cggcggggtg
     4021 cttcacgtac gccttggagc cgtacatgaa ctggggggac aggatgtccc aggcgaaggg
     4081 cagggggccg cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc
     4141 ctcgccctcg ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgcgcacctt
     4201 gaagcgcatg aactctttga tgacctcctc gcccttgctc accatggtgg cgaattctcc
     4261 aggcgatctg acggttcact aaacgagctc tgcttatata ggcctcccac cgtacacgcc
     4321 tacctcgaca tacgttctct atcactgata gggagtaaac tcgacatacg ttctctatca
     4381 ctgataggga taaactcgac atacgttctc tatcactgat agggagtaaa ctcgacatac
     4441 gttctctatc actgataggg agtaaactcg acatacgttc tctatcactg atagggagta
     4501 aactcgacat cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
     4561 gatagggagt aaaccaattc gagatcttcc tagagcatgg ctacgtagat aagtagcatg
     4621 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     4681 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4741 gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag gcccgcaccg
     4801 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4861 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4921 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4981 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5041 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5101 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5161 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5221 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5281 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5341 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5401 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5461 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5521 aaaagatgct gaa
//
