LOCUS       sequence-116996-        6116 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Used for making transgenic mice. Contains a loxP-flanked cassette 
            with a PGK-neo selectable marker and a tpA transcriptional stop 
            sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  AUTHORS   Srinivas S, Watanabe T, Lin CS, William CM, Tanabe Y, Jessell TM, 
            Costantini F
  TITLE     Cre reporter strains produced by targeted insertion of EYFP and ECFP
            into the ROSA26 locus.
  JOURNAL   BMC Dev Biol. 2001  . 1():4.
  PUBMED    11299042
REFERENCE   2  (bases 1 to 6116)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Dev 
            Biol. 2001  . 1():4."
COMMENT     Sequence Label: pBigT
FEATURES             Location/Qualifiers
     source          1..6116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    332..556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     545..562
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     599..615
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     600..616
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(651..684)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    713..847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    967..1101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1221..1355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1503..1964
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     CDS             complement(1974..2777)
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2085..2104)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2695..2714
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2791..3290)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2839..2858)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     3232..3255
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(3320..3353)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3367..3383)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3367..3383)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(3416..3453,3454..3519)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     primer_bind     complement(3546..3565)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3547..3565)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3572..3589)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3572..3588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3581..3603)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3729..4184)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3866..3887)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4078..4097
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4211..4315
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4316..4384,4385..5176)
                     /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(4534..4553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5347..5935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5836..5855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6089..6106
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
      241 aaattaaccc tcactaaagg gaacaaaagc tggtaccgtt ggcgcgccaa ctagaggatc
      301 gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc
      361 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
      421 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
      481 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
      541 caactagaag gcacagtcga ggctgatcag cgagctcgcg gccgcccgcg ggggcccctc
      601 gaggtcgacg gtatcgataa gctagcttgg gctgcaggtc gagggaccta ataacttcgt
      661 atagcataca ttatacgaag ttatattaag ggttccggat cagcttgatg gggatccaga
      721 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      781 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      841 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      901 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
      961 gtcgcagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1021 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1081 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1141 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     1201 tatgatcctc tagagtcgca gatccagaca tgataagata cattgatgag tttggacaaa
     1261 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1321 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1381 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1441 gtggtatggc tgattatgat cctctagagt cgcagatcct ctagagtcgc agatctgcaa
     1501 gcttctgatg gaattagaac ttggcaaaac aatactgaga atgaagtgta tgtggaacag
     1561 aggctgctga tctcgttctt caggctatga aactgacaca tttggaaacc acagtactta
     1621 gaaccacaaa gtgggaatca agagaaaaac aatgatccca cgagagatct atagatctat
     1681 agatcatgag tgggaggaat gagctggccc ttaatttggt tttgcttgtt taaattatga
     1741 tatccaacta tgaaacatta tcataaagca atagtaaaga gccttcagta aagagcaggc
     1801 atttatctaa tcccacccca cccccacccc cgtagctcca atccttccat tcaaaatgta
     1861 ggtactctgt tctcaccctt cttaacaaag tatgacagga aaaacttcca ttttagtgga
     1921 catctttatt gtttaataga tcatcaattt ctgcagactt acagcggatc ccctcagaag
     1981 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     2041 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     2101 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     2161 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     2221 tcatcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     2281 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     2341 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     2401 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     2461 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     2521 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     2581 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     2641 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     2701 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     2761 tcaatggccg atcccatatt ggctgcaggt cgaaaggccc ggagatgagg aagaggagaa
     2821 cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg
     2881 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag
     2941 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac
     3001 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc
     3061 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg
     3121 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt
     3181 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc ccagcggggc
     3241 tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc ggtagaattc
     3301 ctgcaggtcg agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg
     3361 gttccggatc cactagttct agagcggcct cgactctacg ataccgtcga tccccactgg
     3421 aaagaccgcg aagagtttgt cctcaaccgc gagctgtgga aaaaaaaggg acaggataag
     3481 tatgacatca tcaaggaaac cctggactac tgcgccctac agatccctta attaaggcca
     3541 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     3601 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3661 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3721 gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3781 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3841 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3901 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3961 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4021 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4081 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcagtca cgctgcgcgt
     4141 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     4201 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4261 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4501 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4621 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4861 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4921 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5161 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6001 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6061 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
