LOCUS       sequence-340742-        4793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express zebrafish Csf3a, mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  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 4793)
  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-zfCSF3a
FEATURES             Location/Qualifiers
     source          1..4793
                     /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(802..818)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..818)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(821..840)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(822..840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    845..979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(869..888)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     923..942
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1092..1112)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1092..1110)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1131..1147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1131..1147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1155..1171
                     /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(1179..1185,1186..1203,1204..1209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1362..1379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1533..2121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1613..1632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2292..3083,3084..3152))
                     /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     2915..2934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3153..3257)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3283..3738)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3420..3441)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3632..3651
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3809..4793
                     /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 atcccatcga ttcgaattcc tgcagccctg aacatggggc
      121 tttgttacta ctggagtctg gtgattgtgc tcctctgctt tttgattggt gttgctataa
      181 tgaacttcca ggcagctcag ttttttttca cattagtggg gatcgttgtg tctgctcctg
      241 ttcctcagaa actggacatc atgaacaagg atactattga acaagcacac agtttaatca
      301 gcaaaactct ggaagatatt cctgcaactc atgcagcatg ggtaaagagc aagagtctgg
      361 cttggggcag cagcacagat aagctgcagc acttgaaaca ttacataccc tcagcccctg
      421 ttctccagaa catcactgat atcagcagtc tggaaacctg tcttgataaa atagtgaggg
      481 gcctgcaact acatctgaac ctcctgaaag atctcattga agcaacgacc ctgtcgcaga
      541 ctgaccaggt gacagaactg caggctgaca ttcaagaact tgtgcttctt atcgaggagc
      601 ttcagaatca gtcaggattt aacccatcac agcagacatc agaagagcag tcccagagct
      661 tcaagctcaa tctgacccaa cacctgaaaa gtgatttcca gactgaggcc gcggctcacc
      721 taatcctgca tcaactgcgg gatttcagct gtgacatcct tcagaggatc ctcagtattc
      781 gtgtatgaat cgatccgggg ggatccacta gttctagaag ccctatagtg agtcgtatta
      841 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      901 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      961 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1021 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
     1081 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     1141 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     1201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2221 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2281 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2341 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2401 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2461 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2521 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2581 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2641 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2701 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2761 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2821 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2881 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2941 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3001 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3061 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3121 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3181 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3241 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     3301 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3361 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3421 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3481 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3541 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3601 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3661 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3721 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     3781 cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga
     3841 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
     3901 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
     3961 accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg
     4021 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
     4081 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
     4141 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
     4201 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
     4261 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
     4321 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
     4381 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
     4441 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
     4501 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
     4561 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
     4621 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
     4681 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
     4741 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
