LOCUS       sequence-111877-        6745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for iCre mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  AUTHORS   Ohtsuka M, Miura H, Hayashi H, Nakaoka H, Kimura M, Sato M, 
            Gurumurthy CB, Inoko H
  TITLE     Improvement of pronuclear injection-based targeted transgenesis 
            (PITT) by iCre mRNA-mediated site-specific recombination.
  JOURNAL   Transgenic Res. 2013 Aug;22(4):873-5. doi: 
            10.1007/s11248-013-9703-x. Epub 2013 Mar 26.
  PUBMED    23529203
REFERENCE   2  (bases 1 to 6745)
  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: 
            "10.1007/s11248-013-9703-x"; journalName: "Transgenic Res"; date: 
            "2013-08"; volume: "22"; issue: "4"; pages: "873-5"
COMMENT     Sequence Label: pBBI
FEATURES             Location/Qualifiers
     source          1..6745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      949..958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             961..981
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2248..2277
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2293..2310
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2333..2350)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2339..2563
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2609..3037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2696..2715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2906..2927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3046..3066)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3051..3381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3232..3367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3294..3313
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3448..4242
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3502..3521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4112..4131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4418..4539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4455..4474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4509..4528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4588..4604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4588..4604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4601..4623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4612..4628
                     /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(4636..4642,4643..4660,4661..4666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4681..4702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4819..4836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4990..5578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5070..5089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5749..6540,6541..6609))
                     /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     6372..6391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6610..6714)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatggtg
      961 cccaagaaga agaggaaagt ctccaacctg ctgactgtgc accaaaacct gcctgccctc
     1021 cctgtggatg ccacctctga tgaagtcagg aagaacctga tggacatgtt cagggacagg
     1081 caggccttct ctgaacacac ctggaagatg ctcctgtctg tgtgcagatc ctgggctgcc
     1141 tggtgcaagc tgaacaacag gaaatggttc cctgctgaac ctgaggatgt gagggactac
     1201 ctcctgtacc tgcaagccag aggcctggct gtgaagacca tccaacagca cctgggccag
     1261 ctcaacatgc tgcacaggag atctggcctg cctcgccctt ctgactccaa tgctgtgtcc
     1321 ctggtgatga ggagaatcag aaaggagaat gtggatgctg gggagagagc caagcaggcc
     1381 ctggcctttg aacgcactga ctttgaccaa gtcagatccc tgatggagaa ctctgacaga
     1441 tgccaggaca tcaggaacct ggccttcctg ggcattgcct acaacaccct gctgcgcatt
     1501 gccgaaattg ccagaatcag agtgaaggac atctcccgca ccgatggtgg gagaatgctg
     1561 atccacattg gcaggaccaa gaccctggtg tccacagctg gtgtggagaa ggccctgtcc
     1621 ctgggggtta ccaagctggt ggagagatgg atctctgtgt ctggtgtggc tgatgacccc
     1681 aacaactacc tgttctgccg ggtcagaaag aatggtgtgg ctgccccttc tgccacctcc
     1741 caactgtcca cccgggccct ggaagggatc tttgaggcca cccaccgcct gatctatggt
     1801 gccaaggatg actctgggca gagatacctg gcctggtctg gccactctgc cagagtgggt
     1861 gctgccaggg acatggccag ggctggtgtg tccatccctg aaatcatgca ggctggtggc
     1921 tggaccaatg tgaacattgt gatgaactac atcagaaacc tggactctga gactggggcc
     1981 atggtgaggc tgctcgagga tggggactga gaattctgca gatatccagc acagtggcgg
     2041 ccgctcgact atgggcacca aagaacctgt aaacgttatc tttttaaatt gaatgtgcac
     2101 aaataaaagt ttggaaaaga aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2161 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aatacgcatc atacgctgtg
     2221 gcgtacctcg agtctagagg gcccttcgaa caaaaactca tctcagaaga ggatctgaat
     2281 atgcataccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     2341 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2401 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2461 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2521 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2581 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2641 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2701 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2761 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2821 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2881 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2941 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     3001 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3061 tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc
     3121 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3181 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3241 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3301 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3361 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3421 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3481 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3541 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3601 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3661 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3721 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3781 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3841 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3901 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3961 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4021 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4081 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4141 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4201 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4261 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4321 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4381 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4441 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4501 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4561 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4621 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4681 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4741 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4801 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4861 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5161 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     5221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5401 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5521 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5641 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5881 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6721 tccccgaaaa gtgccacctg acgtc
//
