LOCUS       sequence-344913-        4476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for optimised iCre RNA synthesis using T3 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4476)
  AUTHORS   Tromp A, Wang H, Hall TE, Mowry B, Giacomotto J
  TITLE     Optimising the zebrafish Cre/Lox toolbox. Codon improved iCre, new 
            gateway tools, Cre protein and guidelines.
  JOURNAL   Front Physiol. 2023 Aug 2;14:1221310. doi: 
            10.3389/fphys.2023.1221310. eCollection 2023.
  PUBMED    37601640
REFERENCE   2  (bases 1 to 4476)
  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: "Front 
            Physiol."; date: "2023-08-2"; volume: "14:1221310. doi"; pages: " 
            10.3389/fphys.2023.1221310. eCollection 2023"
COMMENT     Sequence Label: 141_pT3TS_iCre
FEATURES             Location/Qualifiers
     source          1..4476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           44..86
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    93..117
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      120..129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             132..152
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1183..1207)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1209..1343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1233..1252)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1287..1306
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1406..1426)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1451..1576
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1664..1683)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1665..1683)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1690..1707)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1690..1706)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1699..1721)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1848..2303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1985..2006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2197..2216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2330..2434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2435..2503,2504..3295)
                     /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     complement(2653..2672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3466..4054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3955..3974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4208..4225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4342..4363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4378..4383,4384..4401,4402..4408)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4416..4432
                     /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)."
     primer_bind     4421..4443
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4440..4456
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4440..4456
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctcaagttt gtacaaaaaa gcaggctccg
      121 ccaccatggt gcccaagaag aagaggaaag tctccaacct gctgactgtg caccaaaacc
      181 tgcctgccct ccctgtggat gccacctctg atgaagtcag gaagaacctg atggacatgt
      241 tcagggacag gcaggccttc tctgaacaca cctggaagat gctcctgtct gtgtgcagat
      301 cctgggctgc ctggtgcaag ctgaacaaca ggaaatggtt ccctgctgaa cctgaggatg
      361 tgagggacta cctcctgtac ctgcaagcca gaggcctggc tgtgaagacc atccaacagc
      421 acctgggcca gctcaacatg ctgcacagga gatctggcct gcctcgccct tctgactcca
      481 atgctgtgtc cctggtgatg aggagaatca gaaaggagaa tgtggatgct ggggagagag
      541 ccaagcaggc cctggccttt gaacgcactg actttgacca agtcagatcc ctgatggaga
      601 actctgacag atgccaggac atcaggaacc tggccttcct gggcattgcc tacaacaccc
      661 tgctgcgcat tgccgaaatt gccagaatca gagtgaagga catctcccgc accgatggtg
      721 ggagaatgct gatccacatt ggcaggacca agaccctggt gtccacagct ggtgtggaga
      781 aggccctgtc cctgggggtt accaagctgg tggagagatg gatctctgtg tctggtgtgg
      841 ctgatgaccc caacaactac ctgttctgcc gggtcagaaa gaatggtgtg gctgcccctt
      901 ctgccacctc ccaactgtcc acccgggccc tggaagggat ctttgaggcc acccaccgcc
      961 tgatctatgg tgccaaggat gactctgggc agagatacct ggcctggtct ggccactctg
     1021 ccagagtggg tgctgccagg gacatggcca gggctggtgt gtccatccct gaaatcatgc
     1081 aggctggtgg ctggaccaat gtgaacattg tgatgaacta catcagaaac ctggactctg
     1141 agactggggc catggtgagg ctgctcgagg atggggactg acacccagct ttcttgtaca
     1201 aagtggggga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1261 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1321 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1381 agggggaggt gtgggaggtt ttttccaact ttattataca tagttgctcg agcggatctg
     1441 gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac ataataccaa
     1501 cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc tcctaataaa
     1561 aagaaagttt cttcacattc taaaaaaaaa aaaaaaaaaa aaaaaaaaaa nncccccccc
     1621 cccccctgca gggccaagtc ggcctctaga ggtacccaat tcgccctata gtgagtcgta
     1681 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     1741 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     1801 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcaaa ttgtaagcgt
     1861 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1921 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     1981 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2041 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2101 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2161 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2221 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2281 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2341 tatttgttta tttttctaaa tacattcaaa tatgtatctg ctcatgagac aataaccctg
     2401 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2461 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2521 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2581 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2641 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2701 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2761 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2821 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2881 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2941 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3001 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3061 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3121 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3181 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3241 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3301 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3361 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3421 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3481 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3541 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3601 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4081 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4141 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4201 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     4261 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4321 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     4381 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4441 aggaaacagc tatgaccatg attacgccaa gctcga
//
