LOCUS       sequence-448147-        5121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of Cre recombinase codon modified for zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5121)
  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 5121)
  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-Cre.zf1
FEATURES             Location/Qualifiers
     source          1..5121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    124..258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(148..167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     202..221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(371..391)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(371..389)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(410..426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(410..426)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(423..445)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    434..450
                     /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(458..464,465..482,483..488))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    503..524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(641..658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(812..1400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(892..911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1571..2362,2363..2431))
                     /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     2194..2213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2432..2536)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2562..3017)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2699..2720)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2911..2930
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3088..4072
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        4107..4125
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4107..4124
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tgtgatgaac tacatcagaa acctggactc tgagacagga gctatggtga gactgctgga
       61 ggatggagac taaaaattca aggcctctcg agcctctaga actatagtga gtcgtattac
      121 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
      181 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      241 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
      301 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
      361 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
      421 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
      481 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
      541 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1441 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1501 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1561 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1621 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1681 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     1741 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1801 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1861 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1921 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1981 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2041 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2101 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2161 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2221 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2281 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2341 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2401 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2461 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     2581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2641 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2761 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3001 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3061 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
     3121 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     3181 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     3241 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
     3301 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     3361 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     3421 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     3481 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     3541 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     3601 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     3661 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     3721 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     3781 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     3841 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
     3901 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     3961 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     4021 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc
     4081 tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt
     4141 tctttttgca ggatctgcca ccatgtccaa tcttctaacc gtccaccaaa atctacctgc
     4201 tctgcctgtg gatgctacat ctgatgaggt gagaaagaac ctgatggaca tgttcagaga
     4261 cagacaggct ttctctgagc acacatggaa gatgctgctg tctgtctgca ggagctgggc
     4321 tgcttggtgt aagctgaaca acagaaagtg gttccctgct gagcctgagg atgtgagaga
     4381 ctacctgctg tacctccagg ctagaggact ggctgtgaag acaatccagc agcacctggg
     4441 acagctgaac atgctgcaca gaaggtctgg actgcctaga ccttctgaca gcaatgctgt
     4501 gagcctggtg atgagaagaa tcagaaagga gaatgtggat gctggagaga gagctaagca
     4561 ggctctggct tttgagagaa cagactttga ccaggtgaga agcctgatgg agaactctga
     4621 cagatgtcag gacatcagaa acctggcttt cctgggaatt gcttacaaca cactgctgag
     4681 aattgctgag attgctagaa tcagagtgaa ggacatcagc agaacagatg gaggaagaat
     4741 gctgatccac attggaagaa caaagacact ggtgagcaca gctggagtgg agaaggctct
     4801 gagcctggga gtgacaaagc tggtggagag atggatctct gtgtctggag tggctgatga
     4861 ccctaacaac tacctgttct gtagagtgag aaagaatgga gtggctgctc cttctgctac
     4921 aagccagctg agcacaaggg ctctggaggg aatctttgag gctacacaca gactgatcta
     4981 tggagctaag gatgactctg gacagagata cctggcttgg tctggacact ctgctagagt
     5041 gggagctgct agagacatgg ctagagctgg agtgagcatc cctgagatca tgcaggctgg
     5101 aggatggaca aatgtgaaca t
//
