LOCUS       sequence-401045-        6232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SPAX8-related truncated mutation (K41*) containing the amino acids 1
            to 40 of human NKX6-2, fused with the Venus fluorescent protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6232)
  AUTHORS   Ferreira-Peralta P, Franca B, Murtinheira F, Rodrigues MS, Herrera F
  TITLE     Is spastic ataxia 8 a protein misfolding disorder?
  JOURNAL   Biochim Biophys Acta Mol Basis Dis. 2023 Sep 9;1870(1):166882. doi: 
            10.1016/j.bbadis.2023.166882.
  PUBMED    37696463
REFERENCE   2  (bases 1 to 6232)
  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: "Biochim 
            Biophys Acta Mol Basis Dis."; date: "2023-09-9"; volume: 
            "1870(1):166882. doi"; pages: " 10.1016/j.bbadis.2023.166882"
COMMENT     Sequence Label: pcDNA3.1 NKX6-2(1-40)-Venus
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1375..1394)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1823..1840)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1829..2053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2099..2527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2186..2205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2396..2417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2536..2556)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2541..2870
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2721..2856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2783..2802
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2937..3731
                     /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(2991..3010)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3601..3620
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3905..4026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3942..3961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3996..4015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4075..4091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4075..4091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4088..4110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4099..4115
                     /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(4123..4129,4130..4147,4148..4153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4168..4189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4306..4323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4477..5065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4557..4576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5236..6027,6028..6096))
                     /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     5859..5878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6097..6201)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gacaccatgg acacaaacag acctggcgcc tttgtgctgt ctagcgctcc tctggccgct
      961 ctgcacaata tggccgagat gaagaccagc ctgtttccat acgctctgca gggacctgcc
     1021 ggctttaagc ttggcggagg tggaagcggc ggaggcggat ctggtaccat ggtttctaag
     1081 ggcgaagaac tgttcaccgg cgtggtgcct atcctggtgg aactggatgg ggatgtgaac
     1141 ggccacaagt tctctgtgtc tggcgaaggc gaaggggatg ccacatacgg caagctgacc
     1201 ctgaagctga tctgcaccac aggcaaactg cccgtgcctt ggcctacact ggtcacaaca
     1261 ctcggctacg gcctgcagtg ctttgccaga tatcccgacc acatgaagca gcacgatttc
     1321 ttcaagagcg ccatgcctga gggctacgtg caagagcgga ccatcttctt caaggacgac
     1381 ggcaactaca agaccagggc cgaagtgaag ttcgagggcg acaccctggt caacagaatc
     1441 gagctgaagg gcatcgactt taaagaggac ggcaacatcc tgggccacaa gctcgagtac
     1501 aactacaaca gccacaacgt gtacatcacc gccgacaagc agaagaacgg catcaaggcc
     1561 aacttcaaga tccggcacaa catcgaggat ggcggcgtgc agctggccga tcactatcag
     1621 cagaacaccc ctatcggaga tggccccgtc ctgctgcctg ataaccacta cctgagctac
     1681 cagtctgccc tgagcaagga ccccaatgag aagagggatc acatggtgct gctggaattc
     1741 gtgacagccg ccggaatcac actcggcatg gacgagctgt acaaatgagg atcctaaggg
     1801 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1861 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1921 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1981 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2041 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2221 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2461 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2641 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2821 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2881 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2941 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3001 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3061 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3121 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3181 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3241 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3301 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3361 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3421 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3481 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3541 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3601 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3661 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3721 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3781 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3841 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3901 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3961 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4021 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4081 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4141 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4201 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4261 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4321 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4381 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagc ccaacccggt
     4801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5281 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5341 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5401 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5461 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5521 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5581 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5641 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5701 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5761 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5821 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5881 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5941 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6001 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6061 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
