LOCUS       sequence-340748-        4769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express zebrafish thrombopoietin (Tpo), mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4769)
  AUTHORS   Svoboda O, Stachura DL, Machonova O, Pajer P, Brynda J, Zon LI, 
            Traver D, Bartunek P
  TITLE     Dissection of vertebrate hematopoiesis using zebrafish 
            thrombopoietin.
  JOURNAL   Blood. 2014 Jul 10;124(2):220-8. doi: 10.1182/blood-2014-03-564682. 
            Epub 2014 May 28.
  PUBMED    24869937
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-2014-03-564682"; journalName: "Blood"; date: 
            "2014-07-10- 10"; volume: "124"; issue: "2"; pages: "220-8"
COMMENT     Sequence Label: pCS2-zfTpo
FEATURES             Location/Qualifiers
     source          1..4769
                     /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(797..816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(798..816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    821..955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(845..864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     899..918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1068..1088)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1068..1086)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1107..1123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1107..1123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1120..1142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1131..1147
                     /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(1155..1161,1162..1179,1180..1185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1200..1221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1338..1355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1509..2097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1589..1608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2268..3059,3060..3128))
                     /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     2891..2910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3129..3233)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3259..3714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3396..3417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3608..3627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3785..4769
                     /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 ttcgaattcg cccttgaagt ctgaaagcgt
      121 gggggccatg cacatgataa agctttcgcg ttacttcgaa aaagacgctc tccaatggaa
      181 agcaacgtga ctggagattt catgttgtcg ctgggcatgg atttaaatag agtgctggtt
      241 gtagttttca gcatggttgt ctctgaactg tctcacgttc acactagacc aattgacttt
      301 gtgtgtgaca gcgaggccag gagagtgatg aacaaggtga aggatctaca agaagaaaag
      361 gtaatctgca gtggcgtaga cgctctcccc tcttctattc aattaccatg catcagcatt
      421 cacaaagcaa cctgggagag aaaatcagta caggagcgca gggccgagat cctgctgtct
      481 ttggggactc tagcgcagga tgttcggtct gccaggacac tcagccaacc tggctgtgga
      541 ctcactctgc tgaagcgctt ggaacacagt atcaacaact acctgcatgt agtcagactg
      601 ctccacgtgg agcaggggga gcaggaaggg ccagcggtag agaggtgtct gggtcagccc
      661 tcgaaggatt tgggtttggt gctgaagtac tttgggcttc tgctcacagg caagctagag
      721 ttgctcattg cagagatggc aaaatggtgc tagggatccc gaagggcgaa ttcaaggcct
      781 ctcgagcctc tagaagccct atagtgagtc gtattacgta gatccagaca tgataagata
      841 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      901 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      961 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     1021 cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
     1081 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1141 ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa
     1201 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1261 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1321 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1381 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1441 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1501 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1561 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1621 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1681 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1741 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1801 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1861 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1921 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1981 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2041 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2101 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2161 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2221 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2281 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2341 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2401 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2461 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2521 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2581 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2641 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2701 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2761 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2821 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2881 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2941 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3001 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3061 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3121 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3181 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3241 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3301 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3361 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3421 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3481 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3541 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3601 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3661 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     3721 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     3781 gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca atatggtgga
     3841 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
     3901 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
     3961 cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
     4021 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
     4081 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
     4141 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
     4201 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
     4261 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
     4321 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
     4381 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
     4441 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
     4501 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
     4561 aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat
     4621 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
     4681 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     4741 tctatataag caatgctcgt ttagggaac
//
