LOCUS       sequence-422144-        8886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Gli1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8886)
  TITLE     Fernandez-Zapico lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8886)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: hGli1 flag3x
FEATURES             Location/Qualifiers
     source          1..8886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4266..4331
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4342..4964
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4541..4560)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4988..5008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5006..5303
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5357..6151
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5411..5430)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6021..6040
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6680..7268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6760..6779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7439..8230,8231..8299))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8062..8081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8300..8404)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8431..8859
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8518..8537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8728..8749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaagctta tgttcaactc gatgacccca ccaccaatca
      961 gtagctatgg cgagccctgc tgtctccggc ccctccccag tcagggggcc cccagtgtgg
     1021 ggacagaagg actgtctggc ccgcccttct gccaccaagc taacctcatg tccggccccc
     1081 acagttatgg gccagccaga gagaccaaca gctgcaccga gggcccactc ttttcttctc
     1141 cccggagtgc agtcaagttg accaagaagc gggcactgtc catctcacct ctgtcggatg
     1201 ccagcctgga cctgcagacg gttatccgca cctcacccag ctccctcgta gctttcatca
     1261 actcgcgatg cacatctcca ggaggctcct acggtcatct ctccattggc accatgagcc
     1321 catctctggg attcccagcc cagatgaatc accaaaaagg gccctcgcct tcctttgggg
     1381 tccagccttg tggtccccat gactctgccc ggggtgggat gatcccacat cctcagtccc
     1441 ggggaccctt cccaacttgc cagctgaagt ctgagctgga catgctggtt ggcaagtgcc
     1501 gggaggaacc cttggaaggt gatatgtcca gccccaactc cacaggcata caggatcccc
     1561 tgttggggat gctggatggg cgggaggacc tcgagagaga ggagaagcgt gagcctgaat
     1621 ctgtgtatga aactgactgc cgttgggatg gctgcagcca ggaatttgac tcccaagagc
     1681 agctggtgca ccacatcaac agcgagcaca tccacgggga gcggaaggag ttcgtgtgcc
     1741 actggggggg ctgctccagg gagctgaggc ccttcaaagc ccagtacatg ctggtggttc
     1801 acatgcgcag acacactggc gagaagccac acaagtgcac gtttgaaggg tgccggaagt
     1861 catactcacg cctcgaaaac ctgaagacgc acctgcggtc acacacgggt gagaagccat
     1921 acatgtgtga gcacgagggc tgcagtaaag ccttcagcaa tgccagtgac cgagccaagc
     1981 accagaatcg gacccattcc aatgagaagc cgtatgtatg taagctccct ggctgcacca
     2041 aacgctatac agatcctagc tcgctgcgaa aacatgtcaa gacagtgcat ggtcctgacg
     2101 cccatgtgac caaacggcac cgtggggatg gccccctgcc tcgggcacca tccatttcta
     2161 cagtggagcc caagagggag cgggaaggag gtcccatcag ggaggaaagc agactgactg
     2221 tgccagaggg tgccatgaag ccacagccaa gccctggggc ccagtcatcc tgcagcagtg
     2281 accactcccc ggcagggagt gcagccaata cagacagtgg tgtggaaatg actggcaatg
     2341 cagggggcag cactgaagac ctctccagct tggacgaggg accttgcatt gctggcactg
     2401 gtctgtccac tcttcgccgc cttgagaacc tcaggctgga ccagctacat caactccggc
     2461 caatagggac ccggggtctc aaactgccca gcttgtccca caccggtacc actgtgtccc
     2521 gccgcgtggg ccccccagtc tctcttgaac gccgcagcag cagctccagc agcatcagct
     2581 ctgcctatac tgtcagccgc cgctcctccc tggcctctcc tttcccccct ggctccccac
     2641 cagagaatgg agcatcctcc ctgcctggcc ttatgcctgc ccagcactac ctgcttcggg
     2701 caagatatgc ttcagccaga gggggtggta cttcgcccac tgcagcatcc agcctggatc
     2761 ggataggtgg tcttcccatg cctccttgga gaagccgagc cgagtatcca ggatacaacc
     2821 ccaatgcagg ggtcacccgg agggccagtg acccagccca ggctgctgac cgtcctgctc
     2881 cagctagagt ccagaggttc aagagcctgg gctgtgtcca taccccaccc actgtggcag
     2941 ggggaggaca gaactttgat ccttacctcc caacctctgt ctactcacca cagcccccca
     3001 gcatcactga gaatgctgcc atggatgcta gagggctaca ggaagagcca gaagttggga
     3061 cctccatggt gggcagtggt ctgaacccct atatggactt cccacctact gatactctgg
     3121 gatatggggg acctgaaggg gcagcagctg agccttatgg agcgaggggt ccaggctctc
     3181 tgcctcttgg gcctggtcca cccaccaact atggccccaa cccctgtccc cagcaggcct
     3241 catatcctga ccccacccaa gaaacatggg gtgagttccc ttcccactct gggctgtacc
     3301 caggccccaa ggctctaggt ggaacctaca gccagtgtcc tcgacttgaa cattatggac
     3361 aagtgcaagt caagccagaa caggggtgcc cagtggggtc tgactccaca ggactggcac
     3421 cctgcctcaa tgcccacccc agtgaggggc ccccacatcc acagcctctc ttttcccatt
     3481 acccccagcc ctctcctccc caatatctcc agtcaggccc ctatacccag ccaccccctg
     3541 attatcttcc ttcagaaccc aggccttgcc tggactttga ttcccccacc cattccacag
     3601 ggcagctcaa ggctcagctt gtgtgtaatt atgttcaatc tcaacaggag ctactgtggg
     3661 agggtggggg cagggaagat gcccccgccc aggaaccttc ctaccagagt cccaagtttc
     3721 tggggggttc ccaggttagc ccaagccgtg ctaaagctcc agtgaacaca tatggacctg
     3781 gctttggacc caacttgccc aatcacaagt caggttccta tcccacccct tcaccatgcc
     3841 atgaaaattt tgtagtgggg gcaaataggg cttcacatag ggcagcagca ccacctcgac
     3901 ttctgccccc attgcccact tgctatgggc ctctcaaagt gggaggcaca aaccccagct
     3961 gtggtcatcc tgaggtgggc aggctaggag ggggtcctgc cttgtaccct cctcccgaag
     4021 gacaggtatg taaccccctg gactctcttg atcttgacaa cactcagctg gactttgtgg
     4081 ctattctgga tgagccccag gggctgagtc ctcctccttc ccatgatcag cggggcagct
     4141 ctggacatac cccacctccc tctgggcccc ccaacatggc tgtgggcaac atgagtgtct
     4201 tactgagatc cctacctggg gaaacagaat tcctcaactc tagtgcctct agaggatccc
     4261 gggctgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4321 acgatgacaa gtagtgatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     4381 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     4441 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     4501 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     4561 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     4621 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     4681 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     4741 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     4801 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     4861 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     4921 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct
     4981 tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5041 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5101 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5161 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5221 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5281 aagtagtgag gaggcttttt tggaggaatt gatcagcttg ggatctgatc aagagacagg
     5341 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5401 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5461 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5521 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5581 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5641 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5701 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5761 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5821 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5881 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5941 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6001 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6061 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6121 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6181 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6241 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6301 atgctggagt tcttcgccca ccccgggctc gatcccctcg cgagttggtt cagctgctgc
     6361 ctgaggctgg acgacctcgc ggagttctac cggcagtgca aatccgtcgg catccaggaa
     6421 accagcagcg gctatccgcg catccatgcc cccgaactgc aggagtgggg aggcacgatg
     6481 gccgctttgg tcgacccgga cgggacgctc ctgcgcctga tacagaacga attgcttgca
     6541 ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt cactgcgtgg atggagcgct
     6601 ggcgcctgct gcgcgacggc gagctgctca ccacccactc gccaagctgg aaccgtaaaa
     6661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7081 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8401 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     8461 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     8521 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8581 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8641 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8701 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8761 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     8821 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     8881 caattt
//
