LOCUS       sequence-340745-        5018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express zebrafish Kitlgb, mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5018)
  AUTHORS   Oltova J, Svoboda O, Machonova O, Svatonova P, Traver D, Kolar M, 
            Bartunek P
  TITLE     Zebrafish Kit ligands cooperate with erythropoietin to promote 
            erythroid cell expansion.
  JOURNAL   Blood Adv. 2020 Dec 8;4(23):5915-5924. doi: 
            10.1182/bloodadvances.2020001700.
  PUBMED    33259600
REFERENCE   2  (bases 1 to 5018)
  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/bloodadvances"; journalName: "Blood Adv"; date: "2020-12-8-
            8"; volume: "4"; issue: "23"; pages: "5915-5924"
COMMENT     Sequence Label: pCS2-zfKitlgb
FEATURES             Location/Qualifiers
     source          1..5018
                     /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"
     primer_bind     complement(1046..1065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1047..1065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1070..1204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1094..1113)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1148..1167
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1317..1337)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1317..1335)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1356..1372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1356..1372)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1369..1391)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1380..1396
                     /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(1404..1410,1411..1428,1429..1434))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1449..1470
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1587..1604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1758..2346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1838..1857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2517..3308,3309..3377))
                     /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     3140..3159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3378..3482)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3508..3963)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3645..3666)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3857..3876
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4034..5018
                     /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 atccggagac acggctgatt tgtttttaat attcccatgt
      121 tccacatgag ggaggttaag attggagaaa gcatttgtgt attagtgttg cttttttcgg
      181 ggcttgtgac ctgctcaggt gtttttggga gccctttaac ggatgatgtg gccacgttag
      241 ataccctgag tgagaatatt cccagtgact acagaatacc catcaagttc atcacaaagg
      301 atgtgggtgg agcttgctgg ctgcatttga acctgtatcc agtcgagagc agtttaaaaa
      361 aattggccat caagtttggc aaccagtcca ccaataaggc caacatcacc attttcataa
      421 caatgctgca ggattttcgt ttcactctga acagtgacga cctggaggac agaatgcaag
      481 ccttcaagtg ccactacaga cgggaaaagt ggccgactag gcgcttcttt agctatgtta
      541 aatccgttct tacagtggcc ggctccacat atggggatat tcctccatgc acacctcctc
      601 cttgccagac cctcgcagct cctcctttca cgccaggtca aagcagacag cagaatggca
      661 tgaactctgc ggttcatggg ctcctggcgc tgctcatcat ccccagtgtg gctattctag
      721 ttctcactat ccaaatggca ttaggaagaa gaggtcgttg tggagcaaga atgcgtgaag
      781 ttgagcctca cgacagagct gaagaaaaca gagatgagct ccacagcgga gctgcacagg
      841 aggacccggc ttccacctcc gccagtgaac aagacagggc gtggctggat tctttagggt
      901 gtgcagacac agaggtctaa taaaaaaaaa aaaagcaaaa ggaaagtcga cggaaacgcc
      961 agtatgctcc ggagttatga agggaatatc cagaacggga tggatcccat cgattcgaat
     1021 tcaaggcctc tcgagcctct agaagcccta tagtgagtcg tattacgtag atccagacat
     1081 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1141 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1201 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1261 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     1321 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1381 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     1441 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1501 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1561 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1621 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1681 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1741 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1801 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1861 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1921 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1981 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2041 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2101 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2161 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2221 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2281 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2341 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2401 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2461 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2521 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2581 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2641 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2701 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2761 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2821 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2881 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2941 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3001 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3061 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3121 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3181 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3241 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3301 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3361 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3421 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3481 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     3541 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3601 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3661 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3721 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3781 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3841 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3901 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3961 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4021 cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat gccaattcaa
     4081 tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt gccaattcaa
     4141 tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat gccaattcaa
     4201 tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg tgccaagttt
     4261 gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg tgccaataat
     4321 ggcggccata ttggctatat gccaggatca atatataggc aatatccaat atggccctat
     4381 gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca tatagtattc
     4441 catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc ccatatacca
     4501 tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa tggtctctat
     4561 atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg gcgcctcccc
     4621 cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac gtcaatagga
     4681 ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt tctcacgccc
     4741 cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat ctattaatag
     4801 taacttggca agtacattac tattggaagt acgccagggt acattggcag tactcccatt
     4861 gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat tgacgtcaat
     4921 ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt aggcgtgcct
     4981 aatgggaggt ctatataagc aatgctcgtt tagggaac
//
