LOCUS       sequence-493997-        5678 bp    DNA     circular SYN 26-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5678)
  AUTHORS   Soriano P.
  TITLE     Generalized lacZ expression with the ROSA26 Cre reporter strain.
  JOURNAL   Nat Genet. 1999 Jan;21(1):70-1.
  PUBMED    9916792
REFERENCE   2  (bases 1 to 5678)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Genet."; date: "1999-01"; volume: "21(1)"; pages: "70-1"
COMMENT     Sequence Label: PGKneotpAlox2
FEATURES             Location/Qualifiers
     source          1..5678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    707..740
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        770..1269
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(805..828)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1202..1221
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1283..2086
                     /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(1346..1365)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1956..1975
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2096..2557
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     polyA_signal    2705..2839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2959..3093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3213..3347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3376..3409
                     /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(3444..3460)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3445..3461)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3490..3510)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3490..3508)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3529..3545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3529..3545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3542..3564)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3553..3569
                     /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(3577..3583,3584..3601,3602..3607))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3622..3643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3760..3777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3931..4519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4011..4030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4690..5481,5482..5550))
                     /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     5313..5332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5551..5655)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atccggaacc cttaatataa cttcgtataa
      721 tgtatgctat acgaagttat taggtccctc gacctgcagg aattctaccg ggtaggggag
      781 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
      841 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
      901 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
      961 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     1021 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     1081 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     1141 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     1201 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     1261 ggcctttcga cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg
     1321 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1381 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1441 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     1501 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     1561 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     1621 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1681 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1741 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     1801 gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat ctcgtcgtga cccatggcga
     1861 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     1921 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     1981 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2041 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagggg atccgctgta
     2101 agtctgcaga aattgatgat ctattaaaca ataaagatgt ccactaaaat ggaagttttt
     2161 cctgtcatac tttgttaaga agggtgagaa cagagtacct acattttgaa tggaaggatt
     2221 ggagctacgg gggtgggggt ggggtgggat tagataaatg cctgctcttt actgaaggct
     2281 ctttactatt gctttatgat aatgtttcat agttggatat cataatttaa acaagcaaaa
     2341 ccaaattaag ggccagctca ttcctcccac tcatgatcta tagatctata gatctctcgt
     2401 gggatcattg tttttctctt gattcccact ttgtggttct aagtactgtg gtttccaaat
     2461 gtgtcagttt catagcctga agaacgagat cagcagcctc tgttccacat acacttcatt
     2521 ctcagtattg ttttgccaag ttctaattcc atcagaagct tgcagatctg cgactctaga
     2581 ggatctgcga ctctagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2641 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2701 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2761 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2821 atcttatcat gtctggatct gcgactctag aggatcataa tcagccatac cacatttgta
     2881 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2941 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3001 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3061 aaactcatca atgtatctta tcatgtctgg atctgcgact ctagaggatc ataatcagcc
     3121 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3181 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3241 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3301 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc atcaagctga
     3361 tccggaaccc ttaatataac ttcgtataat gtatgctata cgaagttatt aggtccctcg
     3421 acctgcagcc caagctagct tatcgatacc gtcgacctcg agggggggcc cggtacccag
     3481 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     3541 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3601 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3661 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3721 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3781 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3841 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3901 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3961 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4021 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4081 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4141 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4201 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4261 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4321 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4381 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4441 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4501 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4561 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4621 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4741 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4801 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4861 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4921 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4981 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5041 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5101 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5161 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5221 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5281 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5341 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5401 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5461 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5521 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5581 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5641 aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
