LOCUS       pHM039_TOPFlash-        5932 bp DNA     circular SYN 07-AUG-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5932)
  AUTHORS   Princeton Network
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    15..24
                     /label=TCF/LEF binding site
     misc_feature    30..39
                     /label=TCF/LEF binding site
     misc_feature    45..54
                     /label=TCF/LEF binding site
     misc_feature    61..70
                     /label=TCF/LEF binding site
     misc_feature    76..85
                     /label=TCF/LEF binding site
     misc_feature    91..100
                     /label=TCF/LEF binding site
     misc_feature    106..115
                     /label=TCF/LEF binding site
     promoter        156..187
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      243..248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             251..994
                     /codon_start=1
                     /label=rtTA
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             join(995..1000,1001..1120)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2 from EGFP
                     /note="mammalian codon-optimized"
                     /translation="KLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV"
     CDS             1124..1126
                     /codon_start=1
                     /label=Fusion Red
                     /translation=""
     polyA_signal    1167..1288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1454..1496
                     /label=mzw116
     misc_feature    complement(1454..1474)
                     /label=mzw121
     promoter        1491..1820
                     /label=pCMV promoter
                     /label=pCMV
     CDS             1829..2563
                     /codon_start=1
                     /gene="TagBFP"
                     /label=TagBFP
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLNAAATPTCNMRD"
     misc_feature    2630..2877
                     /label=3'_PibbgyBack
     misc_feature    complement(2882..2925)
                     /label=mzw117
                     /label=mzw117(1)
     misc_feature    complement(2882..2903)
                     /label=mzw117
     misc_feature    2904..2928
                     /label=mzw122
     misc_feature    3348..4208
                     /label=ampR
     misc_feature    4363..4982
                     /label=pBBR322 Origin
     misc_feature    complement(5355..5381)
                     /label=mzw123
     misc_feature    5362..5381
                     /label=mzw118
     misc_feature    5382..5392
                     /label=mzw188
     misc_feature    5393..5399
                     /label=mzw188
                     /label=mzw188(1)
     misc_feature    5400..5722
                     /label=5' PiggyBac
     misc_feature    complement(5723..5746)
                     /label=mzw119
     misc_feature    complement(5723..5746)
                     /label=mzw189
     misc_feature    complement(5723..5746)
                     /label=mzw189
                     /label=mzw189(1)
     misc_feature    5737..5746
                     /label=mzw120
     polyA_signal    5767..5815
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     misc_feature    5829..5920
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 cgagctctta cgcgagatca aagggggtaa gatcaaaggg ggtaagatca aaggggcgcg
       61 agatcaaagg gggtaagatc aaagggggta agatcaaagg gggtaagatc aaaggggcgc
      121 gcccgcgtgc tagcccgggc tcgagatcta gactctagag ggtatataat ggaagctcga
      181 attccagctt ggcattccgg tactgttggt aaaaagcttg gcattccggt actgttggta
      241 aagccacccc atgagccggc tggacaagag caaagtgatc aacagcgccc tggaactgct
      301 gaacggcgtg ggcatcgagg gcctgaccac ccggaagctg gcccagaaac tgggcgtgga
      361 acagcccacc ctgtactggc acgtgaagaa caagcgggcc ctgctggacg ccctgcccat
      421 cgagatgctg gaccggcacc acacccacag ctgccctctg gaaggcgaga gctggcagga
      481 cttcctgcgg aacaacgcca agagctacag atgcgccctg ctgagccacc gggacggcgc
      541 caaagtgcac ctgggcacca gacccaccga gaagcagtac gagacactgg aaaaccagct
      601 ggccttcctg tgccagcagg gcttcagcct ggaaaacgcc ctgtacgccc tgagcgccgt
      661 gggccacttc accctgggct gcgtgctgga agaacaggaa caccaggtcg ccaaagagga
      721 aagagagaca cccaccaccg acagcatgcc ccccctgctg aagcaggcca tcgagctgtt
      781 cgacagacag ggcgccgagc ccgccttcct gttcggcctg gaactgatca tctgcggcct
      841 cgagaagcag ctgaagtgcg agagcggcgg acccaccgac gccctggacg acttcgacct
      901 ggacatgctg cccgccgatg ccctggatga ttttgatctg gatatgctcc ctgctgacgc
      961 tctcgatgac ttcgatctcg atatgctgcc aggcaagctt agccatggct tcccgccgga
     1021 ggtggaggag caggatgatg gcacgctgcc catgtcttgt gcccaggaga gcgggatgga
     1081 ccgtcaccct gcagcctgtg cttctgctag gatcaatgtg tgataattct agagtcgggg
     1141 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     1201 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     1261 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     1321 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     1381 aatcgataag gatccgtcga ccgatgccct tgagagcctt caacctcgac caaggcagct
     1441 gtagatctta gccatctgct ttttgcttgt actgtcgacc ctgtggaatg tgtgtcagtt
     1501 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1561 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     1621 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     1681 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     1741 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     1801 aggcctaggc ttttgcaaaa agcttaccat gagcgagctg attaaggaga acatgcacat
     1861 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
     1921 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
     1981 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
     2041 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
     2101 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     2161 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
     2221 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga
     2281 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     2341 cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     2401 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt
     2461 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     2521 gcttaatgcg gccgcaactc ccacctgcaa catgcgtgac taacgcccgc cccacgaccc
     2581 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg atactagtta aaagttttgt
     2641 tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata
     2701 aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta
     2761 ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg
     2821 catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa tctagtatac
     2881 gcgtatgcgg ccgcttaatt aatccatggc gcctgatgcg gtattttctc cttacgcatc
     2941 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3001 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3061 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3121 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3181 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3241 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3301 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3361 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3421 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3481 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3541 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3601 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3661 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3721 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3781 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3841 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3901 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3961 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4021 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4081 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4141 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4201 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4261 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4321 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4381 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4441 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4501 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4561 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4621 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4681 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4741 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4801 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4861 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4921 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4981 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5041 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5101 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5161 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     5221 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     5281 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     5341 aacaatttca cacaggaaac agctatgacc atgattacgc cggggcgcgc cattctagat
     5401 taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat attgctctct
     5461 ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg ccgcttggag
     5521 ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact ataacgaccg
     5581 cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta actcatacga
     5641 taattatatt gttatttcat gttctactta cgtgataact tattatatat atattttctt
     5701 gttatagata tcaactagaa tgctagcatg ggcccagatc tccgaggagg tacttggagc
     5761 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     5821 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
     5881 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt ac
//
