LOCUS       sequence-406198-        8861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for making transgenic fish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8861)
  AUTHORS   Wittamer V, Bertrand JY, Gutschow PW, Traver D
  TITLE     Characterization of the mononuclear phagocyte system in zebrafish.
  JOURNAL   Blood. 2011 Jun 30;117(26):7126-35. doi: 
            10.1182/blood-2010-11-321448. Epub 2011 Mar 15.
  PUBMED    21406720
REFERENCE   2  (bases 1 to 8861)
  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"; doi: 
            "10.1182/blood-2010-11-321448"; journalName: "Blood"; date: 
            "2011-06-30- 30"; volume: "117"; issue: "26"; pages: "7126-35"
COMMENT     Sequence Label: mhc2dab:mCherry
FEATURES             Location/Qualifiers
     source          1..8861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     348..370
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     362..379
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     363..379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     412..431
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        412..430
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(458..469,470..657)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    1096..1230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1133..1152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1187..1206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1232..1248
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1233..1249
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1264..1974)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1542..1561)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1811..1829
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    join(6206..6368,6369..6380)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     CDS             6392..6403
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(6413..6433)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6413..6431)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6475..6491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6475..6491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6488..6510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6499..6515
                     /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(6523..6529,6530..6547,6548..6553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6568..6589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6706..6723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6877..7465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6957..6976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7636..8427,8428..8496))
                     /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     8259..8278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8497..8601)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(8627..8861,1..221))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8764..8785)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
       61 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      121 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      181 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      241 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      301 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      361 ttgtaaaacg acggccagtg agcgcgtagg gataacaggg taatgcgcgc gtaatacgac
      421 tcactatagg gcgaattggg taccgggggt caagaaccag aggtgtaaag tacttgagta
      481 attttacttg attactgtac ttaagtatta tttttgggga tttttacttt acttgagtac
      541 aattaaaaat caatactttt acttttactt aattacattt ttttagaaaa aaaagtactt
      601 tttactcctt acaattttat ttacagtcaa aaagtactta ttttttggag atcacttcat
      661 tctattttcc cttgctatta ccaaaccaat tgaattgcgc tgatgcccag tttaatttaa
      721 atgttattta ttctgcctat gaaaatcgtt ttcacattat atgaaattgg tcagacatgt
      781 tcattggtcc tttggaagtg acgtcatgtc acatctatta ccacaatgca cagcaccttg
      841 acctggaaat tagggaaatt ataacagtca atcagtggaa gaaaatggag gaagtatgtg
      901 attcatcagc agctgcgagc agcacagtcc aaaatcagcc acaggatcaa gagcacccgt
      961 ggccgtatct tcgcagatct gatctagagg atcataatca gccataccac atttgtagag
     1021 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1081 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1141 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1201 ctcatcaatg tatcttatca tgtctgaatc ctcgaggtcg acggtatcga taagcttgat
     1261 atcttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
     1321 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt
     1381 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc
     1441 gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc
     1501 ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg
     1561 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
     1621 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
     1681 ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt
     1741 cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag
     1801 ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     1861 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa
     1921 gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctca ccatggtccc
     1981 atcgaattcc tgcagcccgg gggatccgag tctttgaatg tgtcaaatga agaactttcc
     2041 ctcagtgtct tcttcttgtg gtggatttct aaataatctg accaatcagc ttactcccac
     2101 ataagtgttg ctaggtcaaa aactgctttc atttttctgt tttcacaaga caattactgt
     2161 gagtgcatta acacatgtat aaccatagcg gttgtgtaca ctacacaaaa aatgataaaa
     2221 tgtaagaata tttattagtt gtgtctgttt tctaaattaa gaacatgtgt ttatattaga
     2281 ttagtttagt tatatttttt atagaaacag atgaaactgt ttatagttgt gacagtttgc
     2341 tatatattta tttggtaaca tttttgtaaa ttacaatttt ttattttatt taactaaagt
     2401 cttgaaatgt aatttaattt tttaacatca atatattgaa ataatgtcaa tatatttatt
     2461 tgcaaaaaaa aagattttaa aatttatttt attttattat ttcattttag ttaagcctaa
     2521 tttataattt tattgtattt tatattttat tttattaacg tttagtgtgt tattttattt
     2581 aactttaatt tgatatatta ctttattgtt ttgttttatt tgatttgatt ttactttatt
     2641 ttgctttggt ttttgtttta atttattatt ttttatttta ttcagtcaat atattttatt
     2701 cagtattgtt tgagtattct attagtttta ttctattatt tcaatttatt ttagtttact
     2761 ttagtttatt attttattac atacatttta ttttattaat tgtattaacg tttggtatat
     2821 tattttagtt tactttatta ttttacttta ttttaatgac tgattttatt ttactttatt
     2881 ttgctttgat tttagtttga ttttatttta cttattgttt tattttattt agtcaatata
     2941 ttgttgttca atatatattt aattattgta tatattttat tattgtattt tatgttagtt
     3001 tattttaaat taatttattg ttttacttta ttttgatgtc ttattttatt tttctttatt
     3061 gttttattca tattattagt tatttttatt ttattaattt aatattttat tcagtcaata
     3121 tatttattat atataattta ttcatttatt aataatataa tttctttttg tattttagtt
     3181 tagttaaggc tattatttta tttaattatt ttaattgttg gctttgcttt gttttatgtt
     3241 atttttttta atttcttatt atttatttgt ttataactta ttatttatgt ctgtataata
     3301 ttttcgttaa ttcttcaaat gttttgtttt gttttatttt agttataaaa atgtataaaa
     3361 attttgtttt ttaataaata ttttgtcagt caatatatgt ttattcattt ttttattttt
     3421 ttattttatg atatggtttt cttttagctt attttctttt attattttat tttagattat
     3481 tttataatat taattttatt taagcttatt ttattttaat tattgtttat tttaatttaa
     3541 tatttttatc attttatttt agtttactgt tattattatt attttttttt tattatttat
     3601 atgttttttg tttgttttat tatttatgtc tgtgtggtat tttttttaaa tttccaaaag
     3661 tattgttttg tttagtttta ttttagttta tcttaaaaat ttataaaaaa aacgattttt
     3721 aataaatatt ttgtcaatct ttatgtttat ttaatatttt ttattattta gttttattat
     3781 ttaatttgaa tttttttatt tctttatttt attaagttta gtttattgtt tattttattt
     3841 taatttaatt atatatttta ttttagtgca tcattttagt ttattatttt atttttattt
     3901 tattttattt ttttaatttt ttttatttta ttttgtttgt tttattttag tttagtttaa
     3961 aatttaattt aaaaaaaaat atttttgtca agcaatatat ccattcatta attttccttc
     4021 ggcttagtcc ctatatttca gaacctccaa ctattccagc atatgtttta tgcggtggat
     4081 gcccttccag ccgtaaccct gttctgggaa acatctatac actcgtacat acactaagtt
     4141 tgatttagct cattcaattc acttttagca catgtttgaa ctgtggggga aaccggagca
     4201 cccagaggaa atcaatgcaa acatgaggag aacatgcaac tccacgcaga aatgccaaca
     4261 ggctcagcca ggtgagcgac agtgctaacc actgagccat tttttttcta ttagtttatt
     4321 attttatttg agattatttt taatttattt tatttgtgtt cattatttcc ttttatgtaa
     4381 ttaatttatt ttatttcaca gtaaatataa gtgaccaatg taaaaaaaaa aatgtcagta
     4441 tatatgtgga ataatcttta actaaatatt taactatatt taggtttatt ttatagaaac
     4501 agatgaaact gttaattgta acactgtttg caaaatattt aggtcatatt tttgtcagtt
     4561 cttctaacta aaaatactgt ttaatttaat tctatttatt ttagtttaat taagtttacc
     4621 aaagccttga aatatatctt ttttttattg aatacaaatg atttaataca aatcatattt
     4681 gtttgtaaca cactcaattt aaattatgct aaacagatgt cgttaataat aataaatatg
     4741 ttcatatttc aaataatcat gtctactcat tgcttaaaga tttgtttgta acaattttca
     4801 ctgcccacac agctgacatt gtgattgtga tatgtaaaaa gtttttaatg actaaaaagc
     4861 agacacaata gtttcaattt aattatttat ttctaaaggt ttgattattc agtatcaaac
     4921 aactgtgcaa aaaaaaaaag actgaataaa agatcttaaa gtaccactat ttaaagttgc
     4981 attataaaac cacactcgca ttacctgatt taaaattcaa tgaaaatgca taaatgatca
     5041 aagcatgaaa aaattacaat taaattaagc tttgcaatta agtttaaatt tagcataaat
     5101 ttcaagaccc ttttgtcttt gcttatcata agataaaacc agaacacaaa ataacgtgta
     5161 gaattgtcac aagtcacaaa aataagacat atccggaagc aaaaatgatt taatggaaca
     5221 attaaatagt gcatatatgt gtgtttatgt gactgtttac ttcagaggac aagtggattt
     5281 actggcaaaa aatatcaaca aaagttttgt gaatatattt tatgtgtctt ttagttaagg
     5341 caaataactg aaaatttttt ttttacagga tcaagtttat ggagaagaaa atataattca
     5401 tctaacctaa aagctatgga atctaaatgt cctgagaagg atatagtccc aaacctgtat
     5461 gaatgcctaa taatttgtat ggtaactttg atcatcaaaa ctttcgatta ttattgtcac
     5521 acacacacac acacacacac acacacacac acacacacac acacacacac acacacacac
     5581 actgtctttt tccttatacc accatattgg attgtttaaa aattatatcc tttatattta
     5641 aatgaagggc gaagcagtgg tacagtaggt agtgctgtcg cctcacagca agaaggtcgc
     5701 tgggtcgctg gttcgagcct cagctcagtt ggcgtttctg tgaagagttt gcatgttctt
     5761 cctgcgttcg tgttggtttc ctccaggtac tccggttttc cccacagtcc aaagacatgc
     5821 ggtacgcggc cgccaccgcg gcccatctgg cctgtgtttc agacaccagg gagtctctgc
     5881 tcacgtttcc tgctatttgc agcctctcta tcaagactaa tacacctctt cccgcatcgg
     5941 ctgcctgtga gaggcttttc agcactgcag gattgctttt cagccccaaa agagctaggc
     6001 ttgacactaa caattttgag aatcagcttc tactgaagtt aaatctgagg ttttacaact
     6061 ttgagtagcg tgtactggca ttagattgtc tgtcttatag tttgataatt aaatacaaac
     6121 agttctaaag caggataaaa ccttgtatgc atttcattta atgttttttg agattaaaag
     6181 cttaaacaag aatctctagt tttctttctt gcttttactt ttacttcctt aatactcaag
     6241 tacaatttta atggagtact tttttacttt tactcaagta agattctagc cagatacttt
     6301 tacttttaat tgagtaaaat tttccctaag tacttgtact ttcacttgag taaaattttt
     6361 gagtactttt tacacctctg tcaagaactc ctgggagctc cagcttttgt tccctttagt
     6421 gagggttaat tgcgcgcatt accctgttat ccctacgcgc ttggcgtaat catggtcata
     6481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7741 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     8641 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     8701 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     8761 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     8821 aaaaccgtct atcagggcga tggcccacta cgtgaaccat c
//
