LOCUS       sequence-351376-        4769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express zebrafish Csf3b, mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4769)
  AUTHORS   Stachura DL, Svoboda O, Campbell CA, Espin-Palazon R, Lau RP, Zon 
            LI, Bartunek P, Traver D
  TITLE     The zebrafish granulocyte colony-stimulating factors (Gcsfs): 2 
            paralogous cytokines and their roles in hematopoietic development 
            and maintenance.
  JOURNAL   Blood. 2013 Dec 5;122(24):3918-28. doi: 
            10.1182/blood-2012-12-475392. Epub 2013 Oct 15.
  PUBMED    24128862
REFERENCE   2  (bases 1 to 4769)
  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-2012-12-475392"; journalName: "Blood"; date: 
            "2013-12-5- 5"; volume: "122"; issue: "24"; pages: "3918-28"
COMMENT     Sequence Label: pCS2-zfCSF3b
FEATURES             Location/Qualifiers
     source          1..4769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..991
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1026..1044
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1026..1043
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1788..1807)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1789..1807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1812..1946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1836..1855)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1890..1909
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2059..2079)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2059..2077)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2098..2114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2098..2114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2111..2133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2122..2138
                     /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(2146..2152,2153..2170,2171..2176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2191..2212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2329..2346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2500..3088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2580..2599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3259..4050,4051..4119))
                     /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     3882..3901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4120..4224)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4250..4705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4387..4408)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4599..4618
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt caatatggtg
       61 gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg
      121 gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg
      181 gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg
      241 ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc
      301 atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata
      361 tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat
      421 gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg
      481 ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt
      541 ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac
      601 gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca
      661 ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg
      721 acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg
      781 gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca
      841 atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg
      901 ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga
      961 ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac gtcggagcaa
     1021 gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag gatcccatcg
     1081 attcgaattc gcccttaagt ttccccagtt gaacaatcag gctctttaaa agcctccaga
     1141 tccgcacagc agcacccaac accggacacc gagcaccggc gagccgccat gaagacctgc
     1201 tgcctccttc tgctgcacct ctgtgtgtgc cagctgctgt gtgtgcccct acagccacag
     1261 ctaacacaca cacacactga acacactctg agagccttga gtttggccaa gaagatccta
     1321 aacgacattc cagctgtgca ggagctctgc gcacccaaca ctggtttgag ttcctccact
     1381 gaggagtttt tatccagtga gttccagatc ccgacgttgc cgctgctcaa atctgaggat
     1441 ttaacactgg aggagcgtgt gcagcggatg ctcatcggtc tggagcttca tcacagagtc
     1501 ctgctgaagc tgctggagcc ctgccgccca gagaacctac tgaacgacct cgcagagctg
     1561 cgcacgctgc tcctcctcca ggtgccctct gctggtcctc aggtgttgcc gcacacacac
     1621 actctctcgg ctccactcag ccagttcaag ctgcaggtgc gtgtgcgcgc gacactccgg
     1681 cagctgcgca gcttcatgca ggacgtgttc cgcagcctgc gctgcatcag tgtgtctcaa
     1741 tgctaactgg ataagggcga attcaaggcc tctcgagcct ctagaagccc tatagtgagt
     1801 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
     1861 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1921 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1981 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc
     2041 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     2101 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2161 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2221 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2281 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2341 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2401 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2461 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2521 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2581 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2641 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2701 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2761 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2821 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2881 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2941 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3001 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3061 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3121 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3181 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3241 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3301 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3361 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3421 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3481 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3541 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3601 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3661 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3721 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3781 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3841 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3901 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3961 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4021 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4081 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4141 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4201 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     4261 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4321 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4381 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4441 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4501 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4561 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4621 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4681 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     4741 gggcgatcgg tgcgggcctc ttcgctatt
//
