LOCUS       sequence-101771-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of TagRFPT codon modified for zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4827)
  AUTHORS   Horstick EJ, Jordan DC, Bergeron SA, Tabor KM, Serpe M, Feldman B, 
            Burgess HA
  TITLE     Increased functional protein expression using nucleotide sequence 
            features enriched in highly expressed genes in zebrafish.
  JOURNAL   Nucleic Acids Res. 2015 Jan 27. pii: gkv035.
  PUBMED    25628360
REFERENCE   2  (bases 1 to 4827)
  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: "Nucleic 
            Acids Res. 2015 Jan 27. pii: gkv035."
COMMENT     Sequence Label: pCS2-TagRFPT.zf1
FEATURES             Location/Qualifiers
     source          1..4827
                     /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"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    876..1010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(900..919)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     954..973
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1123..1143)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1123..1141)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1162..1178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1162..1178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1175..1197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1186..1202
                     /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(1210..1216,1217..1234,1235..1240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1255..1276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1393..1410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1644..1663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2323..3114,3115..3183))
                     /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     2946..2965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3184..3288)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3314..3769)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3451..3472)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3663..3682
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3843..4827
                     /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 atccgccacc atggtgagca aaggagagga actgattaaa
      121 gaaaacatgc acatgaagct gtacatggag ggaacagtga acaaccacca cttcaagtgt
      181 acatctgagg gagagggaaa gccttatgag ggaacacaga caatgagaat caaggtggtg
      241 gagggaggac ctctgccttt tgcttttgac atcctggcta ccagcttcat gtatggaagc
      301 agaacattca tcaaccacac acagggaatc cctgacttct tcaagcagag cttccctgag
      361 ggattcacat gggagagagt gacaacatat gaggatggag gagtgctgac agctacacag
      421 gacacaagcc tccaggatgg atgtctgatc tacaatgtga agatcagagg agtgaacttc
      481 cctagcaatg gacctgtgat gcagaagaag acactgggat gggaggctaa cacagagatg
      541 ctgtaccctg ctgatggagg actggaggga agaacagaca tggctctgaa gctggtggga
      601 ggaggacacc tgatctgtaa cttcaagaca acatacagaa gcaagaagcc tgctaagaac
      661 ctgaagatgc ctggagtgta ctatgtggac cacagactgg agagaatcaa ggaggctgac
      721 aaggagacat atgtggagca gcatgaggtg gctgtggcta gatactgtga cctgcctagc
      781 aagctgggac acaagctgaa tggaatggat gagctgtaca agtaaaaatt caaggcctct
      841 cgagcctcta gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg
      901 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      961 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1021 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg
     1081 ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat
     1141 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1201 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
     1261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3361 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3421 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3481 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3541 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3601 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3661 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3721 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     3781 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3841 tcgaccatag ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     3901 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     3961 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggact
     4021 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     4081 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     4141 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     4201 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     4261 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     4321 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     4381 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     4441 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     4501 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     4561 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     4621 gtacattact attggaagga cgccagggta cattggcagt actcccattg acgtcaatgg
     4681 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     4741 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     4801 tatataagca atgctcgttt agggaac
//
