LOCUS       pPig_8X_TOPFlash-tdIRFP 6133 bp ds-DNA     circular     29-JAN-2024
DEFINITION  .
KEYWORDS    "marker:ampR"
FEATURES             Location/Qualifiers
     misc_feature    1..18
                     /label="mzw188"
                     /note="label: mzw188 note: mzw188"
     misc_feature    19..341
                     /label="5'piggyBac*"
                     /note="label: 5'piggyBac* note: 5'piggyBac*"
     misc_feature    complement(342..382)
                     /label="mzw119"
                     /note="label: mzw119 note: mzw119"
     misc_feature    363..419
                     /label="mzw129"
                     /note="label: mzw129 note: mzw129"
     misc_feature    383..419
                     /label="mzw116"
                     /note="label: mzw116 note: mzw116"
     misc_feature    420..445
                     /label="pause site"
                     /note="note: RNA polymerase II transcriptional pause signal from the human α2 globin gene"
     misc_feature    420..457
                     /label="5' Topflash spacer"
     misc_feature    458..699
                     /label="Super 8x TOPFlash Promoter"
                     /note="note: from M50 Super 8x TOPFlash.dna"
     misc_feature    468..578
                     /label="7x TCF/LEF consensus "
     promoter        613..644
                     /label="minP"
                     /note="note: minimal TATA-box promoter with low basal activity"
     regulatory      700..705
                     /label="regulatory region"
                     /note="note: vertebrate consensus sequence for strong initiation of translation (Kozak, 1987) regulatory_class: other: "
     source          700..705
                     /label="source"
                     /note="mol_type: other DNA: organism: synthetic DNA construct"
     misc_feature    706..1704
                     /label="Twist1"
     CDS             706..2652
                     /label="tdIRFP"
                     /note="codon_start: 1 transl_table: 1 translation: MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGSDNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFIGSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKITGFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDINYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGLIVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEESGHGTGSTGSGSSSGTSMAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGSDNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFIGSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKITGFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDINYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGLIVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     misc_feature    1680..2652
                     /label="twist2"
     misc_feature    2653..2715
                     /label="h-t2a #1"
                     /note="label: h-t2a #1"
     misc_feature    2716..3315
                     /label="Puromycin"
                     /note="label: Puromycin note: Puromycin"
     misc_feature    3382..3629
                     /label="3'_PibbgyBack"
                     /note="label: 3'_PibbgyBack note: 3'_PibbgyBack"
     misc_feature    complement(3634..3677)
                     /label="mzw117"
                     /note="label: mzw117 note: mzw117"
     misc_feature    3656..3680
                     /label="mzw122"
                     /note="label: mzw122 note: mzw122"
     misc_feature    4100..4960
                     /label="ampR"
                     /note="label: ampR note: ampR"
     misc_feature    5115..5734
                     /label="pBBR322 Origin"
                     /note="label: pBBR322 Origin note: pBBR322 Origin"
     misc_feature    complement(6107..6133)
                     /label="mzw123"
                     /note="label: mzw123 note: mzw123"
ORIGIN
        1 ggggcgcgcc attctagatt aaccctagaa agatagtctg cgtaaaattg acgcatgcat
       61 tcttgaaata ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca
      121 tctcagtcgc cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga
      181 ttttgaacta taacgaccgc gtgagtcaaa atgacgcatg attatctttt acgtgacttt
      241 taagatttaa ctcatacgat aattatattg ttatttcatg ttctacttac gtgataactt
      301 attatatata tattttcttg ttatagatat caactagaat gctagcatgg gcccagatct
      361 ccgaggggac ccgacaggcc cgatctgctt tttgcttgta ctgtcgaccc tgtggaatga
      421 agtgcaggtg ccagaacatt tctctatcga taggtaccga gctcttacgc gagatcaaag
      481 ggggtaagat caaagggggt aagatcaaag gggcgcgaga tcaaaggggg taagatcaaa
      541 gggggtaaga tcaaaggggg taagatcaaa ggggcgcgcc cgcgtgctag cccgggctcg
      601 agatctagac tctagagggt atataatgga agctcgaatt ccagcttggc attccggtac
      661 tgttggtaaa aagcttggca ttccggtact gttggtaaag ccaccATGGC TGAAGGCAGC
      721 GTGGCCCGAC AGCCAGACCT TTTGACTTGT GACGATGAAC CAATCCACAT ACCGGGGGCA
      781 ATACAACCTC ATGGTCTCCT TCTGGCGCTT GCTGCCGACA TGACTATAGT GGCCGGCTCT
      841 GACAACTTGC CGGAATTGAC CGGACTTGCT ATTGGGGCGT TGATTGGGCG CTCTGCCGCT
      901 GATGTATTTG ATTCCGAGAC ACATAATAGG CTTACTATAG CCCTCGCCGA ACCAGGGGCT
      961 GCCGTCGGCG CTCCTATAAC AGTTGGGTTC ACGATGCGAA AAGATGCTGG GTTCATTGGT
     1021 AGCTGGCATC GCCACGATCA ACTTATCTTC CTTGAGCTTG AACCCCCTCA ACGGGACGTT
     1081 GCGGAACCCC AAGCTTTCTT TAGAAGGACC AATTCAGCCA TAAGGCGCCT TCAGGCCGCA
     1141 GAGACATTGG AGTCCGCGTG TGCGGCAGCA GCGCAGGAAG TACGAAAGAT CACGGGATTT
     1201 GACCGGGTTA TGATTTACAG ATTCGCATCT GATTTCTCCG GGGAAGTCAT CGCGGAGGAT
     1261 CGGTGTGCAG AAGTGGAAAG CAAGCTTGGT TTGCATTACC CCGCATCTAC GGTTCCGGCC
     1321 CAAGCGAGGA GACTGTATAC GATAAACCCA GTGAGGATCA TACCTGACAT AAATTATAGA
     1381 CCGGTTCCCG TTACGCCAGA CCTGAACCCC GTCACAGGCA GGCCAATAGA CTTGTCTTTT
     1441 GCAATCCTGC GGTCAGTCTC ACCTGTTCAC CTCGAGTTTA TGAGGAACAT AGGGATGCAT
     1501 GGGACGATGA GCATCTCAAT CCTGAGAGGT GAACGGCTCT GGGGACTTAT TGTTTGTCAT
     1561 CATCGCACAC CGTATTACGT TGACCTTGAT GGTCGCCAGG CCTGCGAACT CGTAGCTCAA
     1621 GTATTGGCCT GGCAGATCGG TGTTATGGAG GAAAGCGGTC ATGGGACTGG GAGTACAGGT
     1681 AGCGGCAGCT CTAGTGGCAC CTCCATGGCA GAAGGGTCCG TAGCAAGGCA ACCTGACTTG
     1741 TTGACCTGTG ATGATGAACC GATTCACATT CCTGGAGCAA TTCAACCGCA TGGGCTGCTC
     1801 CTTGCTTTGG CAGCGGACAT GACGATCGTC GCCGGCTCCG ATAACCTGCC CGAGTTGACG
     1861 GGCTTGGCGA TAGGAGCCCT GATAGGCCGC TCAGCCGCTG ACGTATTCGA TAGCGAAACG
     1921 CATAACCGGC TTACAATCGC CTTGGCTGAA CCGGGCGCGG CCGTGGGAGC ACCGATTACT
     1981 GTAGGCTTTA CAATGAGAAA AGACGCCGGC TTTATCGGGT CATGGCACCG ACATGACCAG
     2041 CTGATTTTCC TGGAATTGGA GCCCCCGCAG CGGGATGTAG CCGAACCACA GGCCTTCTTC
     2101 CGGCGCACTA ACTCCGCAAT TAGGAGACTG CAGGCAGCTG AGACTTTGGA ATCAGCATGC
     2161 GCGGCAGCTG CACAAGAAGT CCGGAAAATC ACGGGTTTTG ACCGAGTCAT GATCTATAGA
     2221 TTCGCGAGCG ATTTCTCAGG AGAAGTTATT GCGGAAGACC GATGCGCGGA GGTAGAATCT
     2281 AAGCTTGGGT TGCACTACCC CGCCTCCACC GTTCCGGCGC AAGCCAGACG GCTCTATACC
     2341 ATTAATCCGG TGCGGATCAT TCCAGATATA AATTACCGGC CTGTACCTGT GACACCGGAT
     2401 TTGAACCCTG TCACGGGCCG ACCGATAGAC CTCAGCTTCG CTATATTGCG ATCTGTGTCA
     2461 CCGGTCCACC TCGAGTTTAT GAGGAATATA GGCATGCATG GTACAATGTC CATTTCCATT
     2521 CTCCGGGGTG AACGGCTTTG GGGCCTCATC GTTTGTCACC ATCGAACACC GTATTACGTC
     2581 GATCTCGACG GCAGACAGGC ATGTGAGTTG GTCGCTCAGG TACTCGCTTG GCAGATAGGG
     2641 GTAATGGAGG AGGGGAGCGG AGAAGGTCGC GGAAGTTTGT TGACCTGCGG GGACGTGGAA
     2701 GAAAACCCGG GTCCAatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     2761 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     2821 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     2881 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     2941 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3001 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     3061 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3121 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     3181 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3241 ttcaccgtca ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc
     3301 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     3361 ccccatgcat cgatactagt taaaagtttt gttactttat agaagaaatt ttgagttttt
     3421 gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat
     3481 gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata
     3541 cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat
     3601 atgattatct ttctagggtt aatctagtat acgcgtatgc ggccgcttaa ttaatccatg
     3661 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     3721 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3781 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3841 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3901 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3961 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4081 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4561 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4681 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4741 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4801 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4861 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4921 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5881 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5941 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6001 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6061 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6121 ccatgattac gcc
//