LOCUS       673_pGL4.R_8xGLI_v2_minPro_Luc2p_Ren.txt.xdna    6205 bp      DNA     circular   11/6/2025
DEFINITION  
ACCESSION   
VERSION     
SOURCE      
  ORGANISM  
COMMENT     Serial Cloner Genbank Format
COMMENT     SerialCloner_Type=DNA
COMMENT     SerialCloner_Comments=Ligation of : pGL4.R_TRE_minPro 211517.txt.xdna [5893  nt] :
             (KpnI[3285] / MluI[3269]) to PCR(641- pd51-GLI-LucII.xdna) 
            [304  nt] : (MluI[10] / KpnI[318])
COMMENT     SerialCloner_Ends=0,0,,0,
FEATURES             Location/Qualifiers
     primer_bind     complement(1173..1190)
                     /label=L4440 vector, forward primer_L4440
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=L4440 - note="L4440 vector, forward primer" - complement(1173..1190) \ 
     rep_origin      complement(1344..1932)
                     /label=high-copy-number ColE1/pMB1/pBR322/pUC origin of _ori
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=direction=LEFT - label=ori - note="high-copy-number ColE1/pMB1/pBR322/pUC origin of  - replication" - complement(1344..1932) \ 
     primer_bind     complement(1424..1443)
                     /label=pBR322 origin, forward primer_pBR322ori-F
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=pBR322ori-F - note="pBR322 origin, forward primer" - complement(1424..1443) \ 
     CDS             complement(2132..2992)
                     /label=confers resistance to ampicillin, carbenicillin, and_AmpR
                     /SerialCloner_Color=&hFF4500
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - product="beta-lactamase" - label=AmpR - note="confers resistance to ampicillin, carbenicillin, and - related antibiotics" - translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI - ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQND \ 
     polyA_signal    969..1017
                     /label=synthetic polyadenylation signal_polyA_signal
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=note="synthetic polyadenylation signal" - 3097..3145 \ 
     polyA_signal    3097..3145
                     /label=synthetic polyadenylation signal_polyA_signal
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=note="synthetic polyadenylation signal" - 3097..3145 \ 
     misc_feature    3159..3250
                     /label=RNA polymerase II transcriptional pause signal from _pause site
                     /SerialCloner_Color=&hDC143C
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=pause site - note="RNA polymerase II transcriptional pause signal from  - the human alpha-2 globin gene" - 3159..3250 \ 
     primer_bind     3199..3218
                     /label=pGL3 vector, forward primer_RVprimer3
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=RVprimer3 - note="pGL3 vector, forward primer" - 3199..3218 \ 
     promoter        3624..3655
                     /label=minimal TATA-box promoter with low basal activity_minP
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=minP - note="minimal TATA-box promoter with low basal activity" - 3332..3363 \ 
     regulatory      3682..3691
                     /label=vertebrate consensus sequence for strong initiation _regulatory
                     /SerialCloner_Color=&h84A4C0
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=regulatory_class="other" - note="vertebrate consensus sequence for strong initiation  - of translation (Kozak, 1987)" - 3390..3399 \ 
     CDS             3688..5337
                     /label=synthetic luc2 version of the luciferase gene_luciferase
                     /SerialCloner_Color=&hFF4500
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - product="firefly luciferase" - label=luciferase - note="synthetic luc2 version of the luciferase gene" - translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA - HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP - ANDIYNERELL \ 
     CDS             5341..5460
                     /label=human codon-optimized_hPEST
                     /SerialCloner_Color=&hFF4500
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - product="PEST degradation sequence from mouse ornithine  - decarboxylase" - label=hPEST - note="human codon-optimized" - translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV" - 5049..5168 \ 
     polyA_signal    5512..5633
                     /label=SV40 polyadenylation signal_SV40 poly(A) signal
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=SV40 poly(A) signal - note="SV40 polyadenylation signal" - 5220..5341 \ 
     primer_bind     complement(5523..5542)
                     /label=SV40 polyA terminator, reverse primer_EBV-rev
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=EBV-rev - note="SV40 polyA terminator, reverse primer" - complement(5231..5250) \ 
     primer_bind     5577..5596
                     /label=SV40 polyA, reverse primer_SV40pA-R
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=SV40pA-R - note="SV40 polyA, reverse primer" - 5285..5304 \ 
     promoter        5827..6184
                     /label=SV40 enhancer and early promoter_SV40 promoter
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=SV40 promoter - note="SV40 enhancer and early promoter" - 5535..5892 \ 
     primer_bind     complement(5850..5870)
                     /label=SV40 enhancer, reverse primer for pBABE vectors_pBABE 3'
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=pBABE 3' - note="SV40 enhancer, reverse primer for pBABE vectors" - complement(5558..5578) \ 
     rep_origin      6035..6170
                     /label=SV40 origin of replication_SV40 ori
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=SV40 ori - note="SV40 origin of replication" - 5743..5878 \ 
     primer_bind     6097..6116
                     /label=SV40 promoter/origin, forward primer_SV40pro-F
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=SV40pro-F - note="SV40 promoter/origin, forward primer" - 5805..5824 \ 
     misc_feature    10..945
                     /label=RLuc
                     /SerialCloner_Color=&h84A4C0
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
     misc_feature    3324..3353
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3362..3391
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3393..3422
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3431..3460
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3462..3491
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3500..3529
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3531..3560
                     /label=Gli-BS
                     /SerialCloner_Color=&h0000FF
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=False
                     /SerialCloner_Arrow=True
     misc_feature    3303..3311
                     /label=Gli-Bs
                     /SerialCloner_Color=&h84A4C0
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
ORIGIN
        1 agcgccacca tgaccagcaa ggtgtacgac cccgagcaga ggaagaggat gatcaccggc 
       61 ccccagtggt gggccaggtg caagcagatg aacgtgctgg acagcttcat caactactac 
      121 gacagcgaga agcacgccga gaacgccgtg atcttcctgc acggcaacgc cgctagcagc 
      181 tacctgtgga ggcacgtggt gccccacatc gagcccgtgg ccaggtgcat catccccgat 
      241 ctgatcggca tgggcaagag cggcaagagc ggcaacggca gctacaggct gctggaccac 
      301 tacaagtacc tgaccgcctg gttcgagctc ctgaacctgc ccaagaagat catcttcgtg 
      361 ggccacgact ggggcgcctg cctggccttc cactacagct acgagcacca ggacaagatc 
      421 aaggccatcg tgcacgccga gagcgtggtg gacgtgatcg agagctggga cgagtggcca 
      481 gacatcgagg aggacatcgc cctgatcaag agcgaggagg gcgagaagat ggtgctggag 
      541 aacaacttct tcgtggagac catgctgccc agcaagatca tgagaaagct ggagcccgag 
      601 gagttcgccg cctacctgga gcccttcaag gagaagggcg aggtgagaag acccaccctg 
      661 agctggccca gagagatccc cctggtgaag ggcggcaagc ccgacgtggt gcagatcgtg 
      721 agaaactaca acgcctacct gagagccagc gacgacctgc ccaagatgtt catcgagagc 
      781 gaccccggct tcttcagcaa cgccatcgtg gagggcgcca agaagttccc caacaccgag 
      841 ttcgtgaagg tgaagggcct gcacttcagc caggaggacg cccccgacga gatgggcaag 
      901 tacatcaaga gcttcgtgga gagagtgctg aagaacgagc agtaaactct agaaccggtc 
      961 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc 
     1021 gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt 
     1081 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg 
     1141 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 
     1201 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 
     1261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 
     1321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 
     1381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 
     1441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 
     1501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 
     1561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 
     1621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 
     1681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 
     1741 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 
     1801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 
     1861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 
     1921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 
     1981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 
     2041 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 
     2101 agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga 
     2161 tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca 
     2221 ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc 
     2281 gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa 
     2341 gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag 
     2401 taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg 
     2461 tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg 
     2521 tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg 
     2581 tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc 
     2641 ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt 
     2701 tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca 
     2761 ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga 
     2821 aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca 
     2881 gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc 
     2941 aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc 
     3001 tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa 
     3061 gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta 
     3121 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac 
     3181 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga 
     3241 acatttctct ggcctaactg gccggtacac gcgTagattc gcgatcgacc tcgacaagca 
     3301 gggaacaccc aagtagaagt tccgacaagc agggaacacc caagtagaag ctctcgacct 
     3361 cgacaagcag ggaacaccca agtagaagct ccgacaagca gggaacaccc aagtagaagc 
     3421 tctcgacctc gacaagcagg gaacacccaa gtagaagctc cgacaagcag ggaacaccca 
     3481 agtagaagct ctcgacctcg acaagcaggg aacacccaag tagaagctcc gacaagcagg 
     3541 gaacacccaa gTAGAAGCTC TCGATCGCGA ATCTGTGgta ccgaggatat caagatctgg 
     3601 cctcggcggc caagcttaga cactagaggg tatataatgg aagctcgact tccagcttgg 
     3661 caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca 
     3721 gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag 
     3781 cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt 
     3841 acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg 
     3901 ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc 
     3961 gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag 
     4021 cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa 
     4081 gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc 
     4141 atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat 
     4201 ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc 
     4261 atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg 
     4321 caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc 
     4381 atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc 
     4441 acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag 
     4501 ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta 
     4561 tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag 
     4621 atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc 
     4681 cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc 
     4741 acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct 
     4801 aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc 
     4861 gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc 
     4921 atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac 
     4981 ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca 
     5041 gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc 
     5101 ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa 
     5161 accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag 
     5221 ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac 
     5281 gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgaat 
     5341 tctcacggct tccctcccga ggtggaggag caggccgccg gcaccctgcc catgagctgc 
     5401 gcccaggaga gcggcatgga tagacaccct gctgcttgcg ccagcgccag gatcaacgtc 
     5461 taaggccgcg actctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca 
     5521 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 
     5581 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca 
     5641 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca 
     5701 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct 
     5761 tccgctgatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagc 
     5821 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc 
     5881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 
     5941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 
     6001 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 
     6061 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 
     6121 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 
     6181 caaaaagctc gattcttctg acact
//