LOCUS       sequence-443492-        6666 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6666)
  AUTHORS   Kaczmarczyk SJ, Green JE
  TITLE     A single vector containing modified cre recombinase and LOX 
            recombination sequences for inducible tissue-specific amplification 
            of gene expression.
  JOURNAL   Nucleic Acids Res 2001 Jun 15;29(12):E56-6.
  PUBMED    11410679
REFERENCE   2  (bases 1 to 6666)
  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: "Nucleic 
            Acids Res"; date: "2001-06-15"; volume: "29(12)"; pages: "E56-6"
COMMENT     Sequence Label: p133 pSV-STOP-luc
FEATURES             Location/Qualifiers
     source          1..6666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    587..635
                     /note="synthetic polyadenylation signal"
     misc_feature    649..740
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     689..708
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     rep_origin      842..977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    complement(1002..1035)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          1569..1634
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1764..1784
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2209..2343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2368..2401)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2431..2440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2437..4089
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2503..2521)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4130..4251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4573..4593
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(4745..4762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4916..5504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4996..5015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5675..6466,6467..6535))
                     /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     6298..6317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6536..6640)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
      541 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
      601 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      661 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      721 aggtgccaga acatttctct atcgataggt accgagctct tacgcgtgct agcccgggct
      781 cgagatctgc gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
      841 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatcgct gactaatttt
      901 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
      961 aggctttttt ggaggcctag tgaacctctt cgagggacct aataacttcg tatagcatac
     1021 attatacgaa gttatattaa gggttccgga tcctcgggga caccaaatat ggcgatctcg
     1081 gccttttcgt ttcttggagc tgggacatgt ttgccatcga tccatctacc accagaacgg
     1141 ccgttagatc tgctgccacc gttgtttcca ccgaagaaac caccgttgcc gtaaccacca
     1201 cgacggttgt tgctaaagaa gctgccaccg ccacggccac cgttgtagcc gccgttgttg
     1261 ttattgtagt tgctactgtt atttctggca cttcttggtt ttcctcttaa gtgaggagga
     1321 acataaccat tctcgttgtt gtcgttgatg cttaaatttt gcacttgttc gctcagttca
     1381 gccataatat gaaatgcttt tcttgttgtt cttacggaat accacttgcc acctatcacc
     1441 acaactaact ttttcccgtt cctccatctc ttttatattt tttttctcga gggatctttg
     1501 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     1561 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
     1621 tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct
     1681 ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg
     1741 ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact
     1801 ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt
     1861 gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa
     1921 aatattctgt aacctttata agtaggcata acagttataa tcataacata ctgttttttc
     1981 ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct
     2041 ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta
     2101 gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2161 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2221 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2281 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     2341 atctgacatg gtaagtaagc tcctaggata acttcgtata gcatacatta tacgaagtta
     2401 taagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa aaacataaag
     2461 aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag
     2521 gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag
     2581 gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa
     2641 cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc
     2701 tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt
     2761 tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt
     2821 tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa
     2881 attattatca tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc
     2941 acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg
     3001 gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc
     3061 gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc
     3121 aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga
     3181 atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt
     3241 gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg
     3301 ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat
     3361 ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc
     3421 aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct
     3481 attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt
     3541 tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc
     3601 gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc
     3661 aacgccttga ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa
     3721 gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag
     3781 gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt
     3841 gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag
     3901 cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc
     3961 gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga
     4021 aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc
     4081 gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     4141 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     4201 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     4261 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     4321 taaaacctct acaaatgtgg taaaatcgat aaggatctga acgatggagc ggagaatggg
     4381 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     4441 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     4501 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     4561 gggactttcc acaccctaac tgacacacat tccacagcgg atccgtcgac cgatgccctt
     4621 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     4681 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     4741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6661 cacctg
//
