LOCUS       sequence-334438-        4790 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 4790)
  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 4790)
  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..4790
                     /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(799..815)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(799..815)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    839..973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(863..882)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     917..936
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1086..1106)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1086..1104)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1125..1141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1125..1141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1138..1160)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1149..1165
                     /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(1173..1179,1180..1197,1198..1203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1218..1239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1356..1373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1527..2115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1607..1626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2286..3077,3078..3146))
                     /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     2909..2928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3147..3251)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3277..3732)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3414..3435)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3626..3645
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3806..4790
                     /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 ggctcagttt tttttcacat tagtggggat cgttgtgtct gctcctgttc
      241 ctcagaaact ggacatcatg aacaaggata ctattgaaca agcacacagc ttaatcagca
      301 aaactctgga agatattcct gcaactcatg cagcatgggt aaagagcaag agtctggctt
      361 ggggcagcag cacagataag ctgcagcact tgaaacatta cataccctca gcccctgttc
      421 tccagaacat cactgatatc agcagtctgg aaacctgtct tgataaaata gtgaggggcc
      481 tgcaactaca tctgaacctc ctgaaagatc tcattgaagc aacgaccctg tcgcagactg
      541 accaggtgac agaactgcag gctgacattc aagaacttgt gcttcttatc gaggagcttc
      601 agaatcagtc aggatttaac ccatcacagc agacatcaga agagcagtcc cagagcttca
      661 agctcaatct gacccaacac ctgaaaagtg atttccagac tgaggccgcg gctcacctaa
      721 tcctgcatca actgcgggat ttcagctgtg acatccttca gaggatcctc agtattcgtg
      781 tatgaatcga tccgggggga tccactagtt ctagaactat agtgagtcgt attacgtaga
      841 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      901 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      961 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1021 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     1081 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     1141 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1201 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1261 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1321 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1381 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1441 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1501 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1561 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1621 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1681 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1741 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1801 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1861 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1921 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     1981 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2041 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2101 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2161 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2221 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2281 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2341 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2401 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2461 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2521 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2581 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2641 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2701 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2761 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2821 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2881 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2941 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3001 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3061 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3121 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3181 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3241 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     3301 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3361 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3421 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3481 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3541 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3601 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3661 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3721 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3781 gggcctcttc gctattacgc cagtcgacca tagccaattc aatatggcgt atatggactc
     3841 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
     3901 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
     3961 atgccaattc aatatggcgg acttggcact gtgccaactg gggaggggtc tacttggcac
     4021 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
     4081 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
     4141 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
     4201 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
     4261 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
     4321 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
     4381 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
     4441 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
     4501 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
     4561 atctattaat agtaacttgg caagtacatt actattggaa ggacgccagg gtacattggc
     4621 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
     4681 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     4741 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
