LOCUS       sequence-203620-       10340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Gli2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10340)
  TITLE     Fernandez-Zapico lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10340)
  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            "Unpublished"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: hGli2 flag3x
FEATURES             Location/Qualifiers
     source          1..10340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5581..5646
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5657..6279
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5856..5875)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6303..6323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6321..6618
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6672..7466
                     /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(6726..6745)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7336..7355
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7994..8582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8074..8093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8753..9544,9545..9613))
                     /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     9376..9395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9614..9718)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9745..10200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9832..9851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10042..10063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(10326..10340,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(10340,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
       61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
      121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      781 attaagcttg cggccgcgaa ttcaatggag acgtctgcct cagccactgc ctccgagaag
      841 caagaagcca aaagtgggat cctggaggcc gctggcttcc ccgacccggg taaaaaggcc
      901 tctcctttgg tggtggctgc agcggcagca gcagcggtag ctgcccaagg agtgccgcag
      961 catctcttgc caccattcca tgcgccccta ccgattgaca tgcgacacca ggaaggaagg
     1021 taccattacg agcctcattc tgtccacggt gtgcacgggc cccctgccct cagcggcagc
     1081 cctgtcatct ctgacatctc cttgatccgg ctttccccgc acccggctgg ccctggggag
     1141 tcccccttca acgcccccca cccgtacgtg aacccccaca tggagcacta cctccgttct
     1201 gtgcacagca gccccacgct ctccatgatc tctgcagcca ggggcctcag ccccgctgat
     1261 gtggcccagg agcaccttaa ggagagggga ctgtttggcc ttcctgctcc aggcaccacc
     1321 ccctcagact attaccacca gatgaccctc gtggcaggcc accccgcgcc ctacggggac
     1381 ctgctgatgc agagcggggg cgctgccagc gcaccccatc tccacgacta cctcaacccc
     1441 gtggacgtgt cccgtttctc cagcccgcgg gtgacgcccc gcctgagccg caagcgggcg
     1501 ctgtccatct ccccactctc agacgccagc ctggacctgc agcggatgat ccgcacctca
     1561 cccaactcgc tagtggccta catcaacaac tcccgaagca gctcagcggc cagcggttcc
     1621 tacgggcatc tgtcagcggg tgccctcagc ccagccttca ccttccccca ccccatcaac
     1681 cccgtggcct accagcagat tctgagccag cagaggggtc tggggtcagc ctttggacac
     1741 acaccacccc tgatccagcc ctcacccacc ttcctggccc agcagcccat ggccctcacc
     1801 tccatcaatg ccacgcccac ccagctcagc agcagcagca actgtctgag tgacaccaac
     1861 cagaacaagc agagcagtga gtcggccgtc agcagcaccg tcaaccctgt cgccattcac
     1921 aagcgcagca aggtcaagac cgagcctgag ggcctgcggc cggcctcccc tctggcgctg
     1981 acgcagggcc aggtgtctgg acacggctca tgtgggtgtg cccttcccct ctcccaggag
     2041 cagctggctg acctcaagga agatctggac agggatgact gtaagcagga ggctgaggtg
     2101 gtcatctatg agaccaactg ccactgggaa gactgcacca aggagtacga cacccaggag
     2161 cagctggtgc atcacatcaa caacgagcac atccacgggg agaagaagga gtttgtgtgc
     2221 cgctggcagg cctgcacgcg ggagcagaag cccttcaagg cgcagtacat gctggtggtg
     2281 cacatgcggc gacacacggg cgagaagccc cacaagtgca cgttcgaggg ctgctcgaag
     2341 gcctactccc gcctggagaa cctgaagaca cacctgcggt cccacaccgg ggagaagcca
     2401 tatgtgtgtg agcacgaggg ctgcaacaaa gccttctcca acgcctcgga ccgcgccaag
     2461 caccagaatc gcacccactc caacgagaaa ccctacatct gcaagatccc aggctgcacc
     2521 aagagataca cagaccccag ctctctccgg aagcatgtga aaacggtcca cggcccagat
     2581 gcccacgtca ccaagaagca gcgcaatgac gtgcacctcc gcacaccgct gctcaaagag
     2641 aatggggaca gtgaggccgg cacggagcct ggcggcccag agagcaccga ggccagcagc
     2701 accagccagg ccgtggagga ctgcctgcac gtcagagcca tcaagaccga gagctccggg
     2761 ctgtgtcagt ccagccccgg ggcccagtcg tcctgcagca gcgagccctc tcctctgggc
     2821 agtgccccca acaatgacag tggcgtggag atgccgggga cggggcccgg gagcctggga
     2881 gacctgacgg cactggatga cacaccccca ggggccgaca cctcagccct ggctgccccc
     2941 tccgctggtg gcctccagct gcgcaaacac atgaccacca tgcaccggtt cgagcagctc
     3001 aagaaggaga agctcaagtc actcaaggat tcctgctcat gggccgggcc gactccacac
     3061 acgcggaaca ccaagctgcc tcccctcccg ggaagtggct ccatcctgga aaacttcagt
     3121 ggcagtgggg gcggcgggcc cgcggggctg ctgccgaacc cgcggctgtc ggagctgtcc
     3181 gcgagcgagg tgaccatgct gagccagctg caggagcgcc gcgacagctc caccagcacg
     3241 gtcagctcgg cctacaccgt gagccgccgc tcctccggca tctcccccta cttctccagc
     3301 cgccgctcca gcgaggcctc gcccctgggc gccggccgcc cgcacaacgc gagctccgct
     3361 gactcctacg accccatctc cacggacgcg tcgcggcgct cgagcgaggc cagccagtgc
     3421 agcggcggct ccgggctgct caacctcacg ccggcgcagc agtacagcct gcgggccaag
     3481 tacgcggcag ccactggcgg ccccccgccc actccgctgc cgggcctgga gcgcatgagc
     3541 ctgcggacca ggctggcgct gctggacgcg cccgagcgca cgctgcccgc cggctgccca
     3601 cgcccactgg ggccgcggcg tggcagcgac gggccgacct atggccacgg ccacgcgggg
     3661 gctgcgcccg ccttccccca cgaggctcca ggcggcggag ccaggcgggc cagcgaccct
     3721 gtgcggcggc ccgatgccct gtccctgccg cgggtgcagc gcttccacag cacccacaac
     3781 gtgaaccccg gcccgctgcc gccctgtgcc gacaggcgag gcctccgcct gcagagccac
     3841 ccgagcaccg acggcggcct ggcccgcggc gcctactcgc cccggccgcc tagcatcagc
     3901 gagaacgtgg cgatggaggc cgtggcggca ggagtggacg gcgcggggcc cgaggccgac
     3961 ctggggctgc cggaggacga cctggtgctt ccagacgacg tggtgcagta catcaaggcg
     4021 cacgccagtg gcgctctgga cgagggcacc gggcaggtgt atcccacgga aagcactggc
     4081 ttctctgaca accccagact acccagcccg gggctgcacg gccagcgcag gatggtggct
     4141 gcggactcca acgtgggccc ctccgcccct atgctgggag gatgccagtt aggctttggg
     4201 gcgccctcca gcctgaacaa aaataacatg cctgtgcagt ggaatgaggt gagctccggc
     4261 accgtagacg ccctggccag ccaggtgaag cctccaccct ttcctcaggg caacctggcg
     4321 gtggtgcagc agaagcctgc ctttggccag tacccgggct acagtccgca aggcctacag
     4381 gctagccctg ggggcctgga cagcacgcag ccacacctgc agccccgcag cggagccccc
     4441 tcccagggca tccccagggt aaactacatg cagcagctgc gacagccagt ggcaggcagc
     4501 cagtgtcctg gcatgactac cactatgagc ccccatgcct gctatggcca agtccacccc
     4561 cagctgagcc ccagcaccat cagtggggcc ctcaaccagt tcccccaatc ctgcagcaac
     4621 atgccagcca agccagggca tctggggcac cctcagcaga cagaagtggc acctgacccc
     4681 accacgatgg gcaatcgcca cagggaactt ggggtccccg attcagccct ggctggagtg
     4741 ccaccacctc acccagtcca gagctaccca cagcagagcc atcacctggc agcctccatg
     4801 agccaggagg gctaccacca ggtccccagc cttctgcctg cccgccagcc tggcttcatg
     4861 gagccccaaa caggcccgat gggggtggct acagcaggct ttggcctagt gcagccccgg
     4921 cctcccctcg agcccagccc cactggccgc caccgtgggg tacgtgctgt gcagcagcag
     4981 ctggcctacg ccagggccac aggccatgcc atggctgcca tgccgtccag tcaggaaaca
     5041 gcagaggctg tgcccaaggg agcgatgggc aacatggggt cggtgcctcc ccagccgcct
     5101 ccgcaggacg caggtggggc cccggaccac agcatgctct actactacgg ccagatccac
     5161 atgtacgaac aggatggagg cctggagaac ctcgggagct gccaggtcat gcggtcccag
     5221 ccaccacagc cacaggcctg tcaggacagc atccagcccc agcccttgcc ctcaccaggg
     5281 gtcaaccagg tgtccagcac tgtggactcc cagctcctgg aggcccccca gattgacttc
     5341 gatgccatca tggatgatgg cgatcactcg agtttgttct cgggtgctct gagccccagc
     5401 ctcctccaca gcctctccca gaactcctcc cgcctcacca ccccccgaaa ctccttgacc
     5461 ctgccctcca tccccgcagg catcagcaac atggctgtcg gggacatgag ctccatgctc
     5521 accagcctcg ccgaggagag caagttcctg aacatgatga cctctagagg atcccgggct
     5581 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     5641 gacaagtagt gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     5701 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     5761 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     5821 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     5881 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     5941 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     6001 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     6061 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     6121 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     6181 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     6241 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     6301 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6361 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     6421 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6481 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6541 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6601 gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag acaggatgag
     6661 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6721 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6781 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6841 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     6901 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6961 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     7021 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     7081 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     7141 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     7201 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     7261 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     7321 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     7381 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     7441 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     7501 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7561 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7621 ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag
     7681 gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag
     7741 cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc
     7801 tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc ttgcaggcat
     7861 ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga gcgctggcgc
     7921 ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg taaaaggccg
     7981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8521 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8581 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8641 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8701 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8821 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8881 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8941 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9001 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9061 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9361 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9721 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     9781 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     9841 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     9901 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     9961 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
    10021 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
    10081 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
    10141 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
    10201 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
    10261 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
    10321 gttttcccag tcacgacgtt
//
