LOCUS       sequence-331153-        4752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express zebrafish Il34, mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4752)
  AUTHORS   Hason M, Mikulasova T, Machonova O, Pombinho AR, van Ham TJ, Irion 
            U, Nusslein-Volhard C, Bartunek P, Svoboda O
  TITLE     M-CSFR/CSF1R signaling regulates myeloid fates in zebrafish via 
            distinct action of its receptors and ligands.
  JOURNAL   Blood Adv. 2022 Jan 3. pii: 483305. doi: 
            10.1182/bloodadvances.2021005459.
  PUBMED    34979548
REFERENCE   2  (bases 1 to 4752)
  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: "Blood Adv. 
            2022 Jan 3. pii: 483305. doi: 10.1182/bloodadvances.2021005459."
COMMENT     Sequence Label: pCS2-zfIl34
FEATURES             Location/Qualifiers
     source          1..4752
                     /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(780..799)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(781..799)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    804..938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(828..847)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     882..901
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1051..1071)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1051..1069)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1090..1106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1090..1106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1103..1125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1114..1130
                     /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(1138..1144,1145..1162,1163..1168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1183..1204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1321..1338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1492..2080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1572..1591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2251..3042,3043..3111))
                     /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     2874..2893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3112..3216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3242..3697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3591..3610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3768..4752
                     /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 atccaccatg gtccagtccg aatgctggct tctcaggggg
      121 ctgctgggtt ttatttgtct attaccagtg tgcagtagtg cagctccaga tctctgtgga
      181 cctctgaaaa cagtccaaga cagtctaaat gcaaccttaa ggagacgtta tatgaaaatg
      241 cactttccca ttaattacac tgtccaagtg cgttatgagg aggtgttcag actgcgaaac
      301 atcagcagac tggtgaatac gagtaatgaa gaagagccag tgctgccgag agatctccag
      361 gatttgtggc tttacgtgag tcaacagggc attaagaagg tcttacgggt actgccagag
      421 aggcatccaa cgcggcgtaa atacctctct gatctggaga atctcttcaa aaaatttgag
      481 acggttttta aagaagggaa tcacgaagac caagagaatg tcagagaacg tccagagagc
      541 ttacaaacca tctgggatca cttgactgaa caagattata aaggatggaa gtcagtcaca
      601 cccaaatcaa tccttgataa ctgttatcgg accatgcttt gcctgtttaa agagtgcttc
      661 acaaaagagg atgacaatta tgattattgt gaagtctata acaggaggaa agaaaggaaa
      721 acaacatagc agcagggatc ccatcgattc gaattcaagg cctctcgagc ctctagaagc
      781 cctatagtga gtcgtattac gtagatccag acatgataag atacattgat gagtttggac
      841 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      901 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      961 ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa
     1021 tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     1081 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1141 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1201 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1261 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3241 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3301 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3361 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3421 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3481 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3541 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3601 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3661 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     3721 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat
     3781 tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat
     3841 tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat
     3901 tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac
     3961 tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa
     4021 gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg
     4081 atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca
     4141 atactatgta ttggccctat gccatatagt attccatata tgggttttcc tattgacgta
     4201 gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca
     4261 tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata
     4321 tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc
     4381 ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat
     4441 ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg
     4501 gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg
     4561 aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc
     4621 gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc
     4681 gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct
     4741 cgtttaggga ac
//
