LOCUS       sequence-190484-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the Distal-less Homeobox 6 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  TITLE     Unpublished Kohtz lab plasmids
REFERENCE   2  (bases 1 to 6314)
  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"
COMMENT     Sequence Label: pcDNA-Dlx6
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     774..794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     868..887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        868..886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1872..1890)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1873..1890)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1910..1927)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1916..2140
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2186..2614
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2273..2292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2483..2504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2623..2643)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2628..2957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2808..2943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2870..2889
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3024..3818
                     /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(3078..3097)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3688..3707
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3992..4113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4029..4048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4083..4102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4162..4178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4162..4178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4175..4197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4186..4202
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4210..4216,4217..4234,4235..4240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4255..4276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4393..4410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4564..5152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4644..4663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5323..6114,6115..6183))
                     /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     5946..5965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6184..6288)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
      901 gtaccgagct cggatccatg atgaccatga ctacgatggc tgacggcttg gaaggccagg
      961 actcgtccaa atccgccttc atggagttcg ggcagcagca acagcagcag cagcaacaac
     1021 agcagcagca acagcagcag cagcagcagc aacagcagcc gccgccgccg ccaccgccgc
     1081 cgccgccgca gccgcactcg cagcagacct ccccggccat ggcaggcgca cattaccctc
     1141 tgcactgctt gcactcggcc gcggcggcgg cggcggcggc cggctcccac catcaccacc
     1201 accagcacca ccaccacggc tcgccctacg cgtcgagcgg aggcaactcc tacaaccacc
     1261 gatcgctcgc cgcctacccc tacatgagcc actcgcagca cagcccttac ctccagtcct
     1321 accacaacag cagcgcggcc gcccagacgc gcggggacga cacagatcaa caaaaaacga
     1381 cagtgatcga aaacggggaa atcaggttca acggaaaggg gaaaaagatt cggaagcctc
     1441 ggaccattta ttccagcctg cagctccagg ctttaaacca tcgctttcag cagactcaat
     1501 acctggccct tcccgagaga gccgaactgg ctgcttcctt aggactgaca caaacacagg
     1561 tgaagatatg gtttcagaat aagcgctcta agtttaagaa attgctgaag cagggtagta
     1621 acccacacga gagtgacccc ctcccgggtt cagcagccct gtcaccacga tcaccagccc
     1681 tgcctccagt gtgggacgtt tctgcctctg ccaagggcgt cagtatgcct cccaacagct
     1741 acatgccggg gtattcacac tggtattcct caccacacca ggacaccatg cagagaccac
     1801 agatgatgta ggaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     1861 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     1921 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1981 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2041 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2101 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2161 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2221 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2281 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2341 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2401 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2461 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2521 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2581 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2641 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2701 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2761 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2821 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2881 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2941 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3001 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3061 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3121 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3181 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3241 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3301 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3361 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3421 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3481 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3541 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3601 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3661 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3721 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3781 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3841 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3901 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3961 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4021 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4081 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4141 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6301 aaaagtgcca cctg
//
