LOCUS       sequence-154507-        4534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning of zebrafish mitfa gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4534)
  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 4534)
  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: pBS-mitfa
FEATURES             Location/Qualifiers
     source          1..4534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2296..2312)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2296..2312)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2349..2367)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2388..2404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2388..2404)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2401..2423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2412..2428
                     /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(2436..2442,2443..2460,2461..2466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2481..2502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2619..2636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2790..3378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2870..2889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3549..4340,4341..4409))
                     /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     4172..4191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4410..4514)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gagttttttt tttttttttt tattcaagct cctctttata tcatccgcga
      721 gaatccttca aaaatcaact aattgtttac acgttttcat tcttgactag actctgacat
      781 gataaatgaa aaaatatata cacaagcaca ctaaatattc tgaaatacaa gattaatact
      841 ttacgaaatt gcgctcttca tgatctggat gtaacactgg tacttacaag acacacacac
      901 acacaaatct aattgtctaa ctttggttca tgaaatttag ttggcattgc taacagccat
      961 tgtcatgttc gtccatactg ctgctgccgc tactgttgga aggtactggg gatccggagg
     1021 acaacagcgg gtcgctgctg cccaccggag acagggtgtt gtccataagc atgtcctcca
     1081 gcttggtgga ggccttgttt gggctgtcgt agacccccgg ctcgccgtcc tccgtgggac
     1141 tgtcattgta gctgatggtg ccgttgttga ggtccagagt ggtaggacgg gacatgtcag
     1201 gactgggaag gtgtgggtac aaattggatg tgcagtcccc catcccaggc tcttgtttta
     1261 ttgctcgagc cactaactca gcggaataaa ggctggaaga agctacaacg gtgagtccat
     1321 gagcacgggc ttgcatttca agttcctgaa tacggagcag gagatgtctg tttgcgtgtt
     1381 ctagtctctt ctgtctgttt tccagctctt ttgctttctg ctgctctttc tgcaatttcc
     1441 taatgtaatc cactgatgct ttcaggatgg tgcctttatt ccacctcatg tctggatcat
     1501 ttgacttggg aattaaagtc cccagctcct taattctgtc attgatatta aaccgccttc
     1561 ttcgttcaat gaggttgtgg ttgtccttct tttgtctctc ttttgccagg gctctgactt
     1621 ctgcttctac tggaaagcca tcaggtcttt gataagagtc aaacttgcca catggtccag
     1681 ctttgtccat aatgtgcatc atgcccggag atggagtaac ggataattcc cttttgacgg
     1741 ccggcaggct tgaggggcag gagttactga tggaaactcc agctggagga agagcatgat
     1801 tgctgtacat gtccagcagg tttgctgaaa ctggaatcgt gtttgtcatt tgaagtcctg
     1861 catccatgaa cccaagaatg tcatcactgt agcttgattc caaacttata atgtcttcaa
     1921 ttacatcgtc catctctttc tcacagttga gggtgagaag ggccataggg ctgttgggag
     1981 cgctggctcc gggtcccggg gtcatgccgt gctcggcggg ctggctgggg ccgggccctg
     2041 tgctggcctg tgggctcagc ttggctccca gtgcactgga caggtagtgc ttcacctgct
     2101 gcctctggct ctgctggatg tggtacttcg agggggtttc caggtgggtc tgaacctggt
     2161 agtgactgta ctcgagcatc tccaacatgt tcaactatgt gttagcttca aaaaagtatt
     2221 ccctttgtga atttaggtga tgtgcaagaa ctgaccagtc gtcttggcct cgtgccgaat
     2281 tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     2341 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
     2401 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2461 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2521 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2581 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4501 ataggggttc cgcgcacatt tccccgaaaa gtgc
//
