LOCUS       sequence-101782-        4725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of bacterial nitroreductase NfsB codon modified for 
            zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4725)
  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 4725)
  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-NfsB.zf1
FEATURES             Location/Qualifiers
     source          1..4725
                     /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    774..908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(798..817)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     852..871
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1021..1041)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1021..1039)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1060..1076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1060..1076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1073..1095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1084..1100
                     /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(1108..1114,1115..1132,1133..1138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1153..1174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1291..1308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1462..2050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1542..1561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2221..3012,3013..3081))
                     /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     2844..2863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3082..3186)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3212..3667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3349..3370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3561..3580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3741..4725
                     /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 atggatatta ttagtgtggc cctgaagaga
      121 catagcacaa aggcttttga tgctagcaag aagctgacac ctgagcaggc tggacagatc
      181 aagacactgc tccagtacag ccctagcagc acaaacagcc agccttggca cttcattgtg
      241 gctagcacag aggagggaaa ggctagagtg gctaagtctg ctgctggaaa ctatgtgttc
      301 aatgagagaa agatgctgga tgctagccat gtggtggtgt tctgtgctaa gacagctatg
      361 gatgatgtgt ggctgaagct ggtggtggac caggaggatg ctgatggaag atttgctaca
      421 cctgaggcta aggctgctaa tgacaaggga agaaagttct ttgctgacat gcacagaaag
      481 gacctgcatg atgatgctga gtggatggct aagcaggtgt acctgaatgt gggaaacttc
      541 ctgctgggag tggctgctct gggactggat gctgtgccta ttgagggatt tgatgctgct
      601 atcctggatg ctgagtttgg actgaaggag aagggataca caagcctggt ggtggtgcct
      661 gtgggacacc actctgtgga ggacttcaat gctacactgc ctaagagcag actgcctcag
      721 aacatcacac tgacagaggt gtgatctaga actatagtga gtcgtattac gtagatccag
      781 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      841 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      901 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
      961 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     1021 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1081 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     1141 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1201 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1261 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1861 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     1921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2221 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2281 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2341 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2401 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2461 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2521 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2581 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2641 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2701 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2761 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2821 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2881 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2941 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3001 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3061 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3121 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3181 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     3241 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3301 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3361 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3421 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3481 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3541 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3601 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3661 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     3721 tcttcgctat tacgccagtc gaccatagcc aattcaatat ggcgtatatg gactcatgcc
     3781 aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc
     3841 aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc
     3901 aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc
     3961 caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc
     4021 caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg
     4081 gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat
     4141 agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca
     4201 tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg
     4261 tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg
     4321 cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc
     4381 aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct
     4441 cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta
     4501 ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca ttggcagtac
     4561 tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga
     4621 cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg
     4681 cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaac
//
