LOCUS       sequence-262625-        4766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses zebrafish granulocyte colony stimulating factor (csf3a).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4766)
  AUTHORS   Rajan V, Collett K, Woodside R, Prykhozhij SV, Moksa M, Carles A, 
            Wong M, Liebman M, Hirst M, Berman JN
  TITLE     Stress hematopoiesis induces a proliferative advantage in TET2 
            deficiency.
  JOURNAL   Leukemia. 2021 Sep 30. pii: 10.1038/s41375-021-01427-7. doi: 
            10.1038/s41375-021-01427-7.
  PUBMED    34588613
REFERENCE   2  (bases 1 to 4766)
  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: "Leukemia. 
            2021 Sep 30. pii: 10.1038/s41375-021-01427-7. doi: 
            10.1038/s41375-021-01427-7."
COMMENT     Sequence Label: pCS2+ gcsf
FEATURES             Location/Qualifiers
     source          1..4766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..984
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1019..1037
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1019..1036
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1802..1936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1826..1845)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1880..1899
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2049..2069)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2049..2067)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2088..2104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2088..2104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2101..2123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2112..2128
                     /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(2136..2142,2143..2160,2161..2166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2181..2202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2319..2336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2490..3078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2570..2589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3249..4040,4041..4109))
                     /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     3872..3891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4110..4214)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4240..4695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4377..4398)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4589..4608
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
       61 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
      121 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
      181 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
      241 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
      301 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
      361 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
      421 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
      481 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
      541 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
      601 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
      661 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
      721 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
      781 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
      841 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
      901 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
      961 ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat
     1021 ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca tcgattcgaa
     1081 ttcgccacca tgatggggct ttgttactac tggagtctgg tgattgtgct cctctgcttt
     1141 ttgattggtg ttgctataat gaacttccag gctcagtttt ttttcacatt agtggggatc
     1201 gttgtgtctg ctcctgttcc tcagaaactg gacatcatga acaaggatac tattgaacaa
     1261 gcacacagtt taatcagcaa aactctggaa gatattcctg caactcatgc agcatgggta
     1321 aagagcaaga gtctggcttg gggcagcagc acagataagc tgcagcactt gaaacattac
     1381 ataccctcag cccctgttct ccagaacatc actgatatca gcagtctgga aacctgtctt
     1441 gataaaatag tgaggggcct gcaactacat ctgaacctcc tgaaagatct cattgaagca
     1501 acgaccctgt cgcagactga ccaggtgaca gaactgcagg ctgacattca agaacttgtg
     1561 cttcttatcg aggagcttca gaatcagtca ggatttaacc catcacagca gacatcagaa
     1621 gagcagtccc agagcttcaa gctcaatctg acccaacacc tgaaaagtga tttccagact
     1681 gaggccgcgg ctcacctaat cctgcatcaa ctgcgggatt tcagctgtga catccttcag
     1741 aggatcctca gtattcgtgt atgactcgag cctctagaac tatagtgagt cgtattacgt
     1801 agatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1861 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1921 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1981 ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc
     2041 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     2101 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag
     2161 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2221 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2281 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2401 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2941 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3001 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3061 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3121 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3181 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3241 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3301 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3361 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3421 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3481 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3541 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3601 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3661 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3721 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3781 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3841 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3901 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3961 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4021 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4081 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4141 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4201 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     4261 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4321 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4381 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4441 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4501 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4561 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4621 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4681 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4741 tgcgggcctc ttcgctatta cgccag
//
