LOCUS       sequence-154510-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of zebrafish mitfa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  AUTHORS   Lister JA, Robertson CP, Lepage T, Johnson SL, Raible DW
  TITLE     nacre encodes a zebrafish microphthalmia-related protein that 
            regulates neural-crest-derived pigment cell fate.
  JOURNAL   Development. 1999 Sep;126(17):3757-67.
  PUBMED    10433906
REFERENCE   2  (bases 1 to 5855)
  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: 
            "Development."; date: "1999-09"; volume: "126(17)"; pages: "3757-67"
COMMENT     Sequence Label: pCS2-MT-mitfa
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             142..171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             181..210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             220..249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             259..288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             322..351
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1904..2038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1928..1947)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1982..2001
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2151..2171)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2151..2169)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2190..2206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2190..2206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2203..2225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2214..2230
                     /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(2238..2244,2245..2262,2263..2268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2283..2304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2421..2438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2592..3180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2672..2691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3351..4142,4143..4211))
                     /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     3974..3993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4212..4316)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4342..4797)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4479..4500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4691..4710
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4871..5855
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggagcaaaag ctcatttctg aagaggactt gaattcaagg
      361 cctctcgagt acagtcacta ccaggttcag acccacctgg aaaccccctc gaagtaccac
      421 atccagcaga gccagaggca gcaggtgaag cactacctgt ccagtgcact gggagccaag
      481 ctgagcccac aggccagcac agggcccggc cccagccagc ccgccgagca cggcatgacc
      541 ccgggacccg gagccagcgc tcccaacagc cctatggccc ttctcaccct caactgtgag
      601 aaagagatgg acgatgtaat tgaagacatt ataagtttgg aatcaagcta cagtgatgac
      661 attcttgggt tcatggatgc aggacttcaa atgacaaaca cgattccagt ttcagcaaac
      721 ctgctggaca tgtacagcaa tcatgctctt cctccagctg gagtttccat cagtaactcc
      781 tgcccctcaa gcctgccggc cgtcaaaagg gaattatccg ttactccatc tccgggcatg
      841 atgcacatta tggacaaagc tggaccatgt ggcaagtttg actcttatca aagacctgat
      901 ggctttccag tagaagcaga agtcagagcc ctggcaaaag agagacaaaa gaaggacaac
      961 cacaacctca ttgaacgaag aaggcggttt aatatcaatg acagaattaa ggagctgggg
     1021 actttaattc ccaagtcaaa tgatccagac atgaggtgga ataaaggcac catcctgaaa
     1081 gcatcagtgg attacattag gaaattgcag aaagagcagc agaaagcaaa agagctggaa
     1141 aacagacaga agagactaga acacgcaaac agacatctcc tgctccgtat tcaggaactt
     1201 gaaatgcaag cccgtgctca tggactcacc gttgtagctt cttccagcct ttattccgct
     1261 gagttagtgg ctcgagcaat aaaacaagag cctgggatgg gggactgcac atccaatttg
     1321 tacccacacc ttcccagtcc tgacatgtcc cgtcctacca ctctggacct caacaacggc
     1381 accatcagct acaatgacag tcccacggag gacggcgagc cgggggtcta cgacagccca
     1441 aacaaggcct ccaccaagct ggaggacatg cttatggaca acaccctgtc tccggtgggc
     1501 agcagcgacc cgctgttgtc ctccggatcc ccagtacctt ccaacagtag cggcagcagc
     1561 agtatggacg aacatgacaa tggctgttag caatgccaac taaatttcat gaaccaaagt
     1621 tagacaatta gatttgtgtg tgtgtgtgtc ttgtaagtac cagtgttaca tccagatcat
     1681 gaagagcgca atttcgtaaa gtattaatct tgtatttcag aatatttagt gtgcttgtgt
     1741 atatattttt tcatttatca tgtcagagtc tagtcaagaa tgaaaacgtg taaacaatta
     1801 gttgattttt gaaggattct cgcggatgat ataaagagga gcttgaataa aaaaaaaaaa
     1861 aaaaaactcg agcctctaga actatagtga gtcgtattac gtagatccag acatgataag
     1921 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1981 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2041 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     2101 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     2161 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2341 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2401 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2461 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2521 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3001 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3241 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3301 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4321 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4381 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4441 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4501 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4561 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4621 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4681 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4741 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     4801 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4861 tacgccagtc gaccatagcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     4921 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     4981 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     5041 ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     5101 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     5161 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     5221 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     5281 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     5341 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     5401 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     5461 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     5521 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     5581 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     5641 cttggcaagt acattactat tggaaggacg ccagggtaca ttggcagtac tcccattgac
     5701 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     5761 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     5821 gggaggtcta tataagcaat gctcgtttag ggaac
//
