pLZ348

LOCUS       pLZ348             8777 bp DNA     circular SYN 14-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8777)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jul 19, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    228
                     /label=E2F-1
     misc_feature    232
                     /label=CMV
     misc_feature    234..245
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    248..259
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    262..273
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    276..287
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    290..301
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    302..306
                     /label=E2F-1
     misc_feature    307..454
                     /label=hcmvmin
     misc_feature    376..390
                     /label=New Feature
     primer_bind     435..455
                     /label=181-WKG-593-F
     misc_feature    477..484
                     /label=New Feature
     misc_feature    487..491
                     /label=NheI
     misc_recomb     496..529
                     /label=LoxP
     protein_bind    complement(496..529)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      569..578
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             578..1276
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    1293..1311
                     /label=linker
     CDS             1313..2011
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    2028..2046
                     /label=linker
     CDS             2048..2746
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    2763..2782
                     /label=linker
     misc_feature    2783..3724
                     /label=NSP3
     misc_feature    2783..2796
                     /label=NSP3
     primer_bind     complement(2794..2813)
                     /label=179-WKG-592-R
     polyA_signal    3753..3977
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      3986..4170
                     /label=SV40 late polyA
     misc_recomb     4207..4240
                     /label=LoxP
     protein_bind    complement(4207..4240)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4247..4295
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    4311..4318
                     /label=Secretory (IgK)
                     /label=nonstandard type:  signal
     misc_feature    4320..4321
                     /label=NheI
     misc_feature    4332
                     /label=New Feature
     misc_feature    4332
                     /label=New Feature
     misc_feature    4337..4342
                     /label=New Feature
     misc_feature    4337..4342
                     /label=New Feature
     terminator      4371..4598
                     /label=bGH polyA
     rep_origin      4644..5072
                     /label=f1 bacteriophage origin of replication
                     /label=f1 bacteriophage origin of replication; arrow _f1 
                     ori
     primer_bind     complement(4658..4682)
                     /label=184-WKG-599-R
     primer_bind     4943..4962
                     /label=189-WKG-612-F
     promoter        5086..5415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     misc_feature    5482..6273
                     /label=Kan/neoR
                     /label=nonstandard type: resistance
     polyA_signal    6450..6571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6616..6636)
                     /label=M13-rev
     primer_bind     complement(6620..6636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(6642..6664)
                     /label=LacO
     protein_bind    6644..6660
                     /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(6668..6698)
                     /label=lac promoter
     protein_bind    6713..6734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6840..7780
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(7781..8641)
                     /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"
     promoter        complement(8642..8746)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtattggat
      241 ccaatgcatt ggatccaatg gattggatcc aatcgattgg atccaattga ttggatccaa
      301 tctagctcga gctcggtacc cgggtcgagt aggcgtgtac ggtgggaggc ctatataagc
      361 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      421 catagaagac accgggaccg atccagcctc cgcggccccg aattcgagct cgcccgggga
      481 tcctcgctag cgcttataac ttcgtatagc atacattata cgaagttatc ggcgcgccgc
      541 ggccgcgaat tcgacccaag ctggctaggc caccatggac atcgagaagc tgtgcaagaa
      601 ggcggaagag gaggccaagg aggcccagga gaaggccgac gagctgagac aacgacaccc
      661 cgattcgcag gcggcggaag acgcggagga tctcgcaaat ctggctgtcg ccgccgtgct
      721 gaccgcttgt ctgctggccc aggagcatcc caacgccgat atcgctaagc tgtgcatcaa
      781 ggccgcgagc gaagctgcgg aggccgcgtc aaaagccgcc gagctggccc aacggcaccc
      841 agacagtcaa gcagccagag atgctatcaa actcgcctct caagccgccc gcgccgtgat
      901 cctggccatc atgctcgccg cagagaatcc caacgctgat attgcaaagc tatgtattaa
      961 ggcggccagc gaagccgcag aggctgcctc gaaagctgcg gaacttgcac agcggcaccc
     1021 cgactcccag gccgcccgag atgcaattaa actggccagt caggccgctg aggcggtgga
     1081 gagagccatc tggctggccg ccgagaaccc caatgccgat atcgccaaaa aatgcatcaa
     1141 agcggcctcc gaagcagccg aggaggcctc taaagctgcc gaagaggctc aaagacaccc
     1201 agacagccag aaggcacgcg acgagatcaa ggaggcctct cagaaggccg aggaggtgaa
     1261 ggagagatgc aagagcgcca gcggaagtgg cggaggaggc gacgtcgcta gtgacatcga
     1321 gaagctgtgc aagaaggcgg aagaggaggc caaggaggcc caggagaagg ccgacgagct
     1381 gagacaacga caccccgatt cgcaggcggc ggaagacgcg gaggatctcg caaatctggc
     1441 tgtcgccgcc gtgctgaccg cttgtctgct ggcccaggag catcccaacg ccgatatcgc
     1501 taagctgtgc atcaaggccg cgagcgaagc tgcggaggcc gcgtcaaaag ccgccgagct
     1561 ggcccaacgg cacccagaca gtcaagcagc cagagatgct atcaaactcg cctctcaagc
     1621 cgcccgcgcc gtgatcctgg ccatcatgct cgccgcagag aatcccaacg ctgatattgc
     1681 aaagctatgt attaaggcgg ccagcgaagc cgcagaggct gcctcgaaag ctgcggaact
     1741 tgcacagcgg caccccgact cccaggccgc ccgagatgca attaaactgg ccagtcaggc
     1801 cgctgaggcg gtggagagag ccatctggct ggccgccgag aaccccaatg ccgatatcgc
     1861 caaaaaatgc atcaaagcgg cctccgaagc agccgaggag gcctctaaag ctgccgaaga
     1921 ggctcaaaga cacccagaca gccagaaggc acgcgacgag atcaaggagg cctctcagaa
     1981 ggccgaggag gtgaaggaga gatgcaagag cgccagcgga agtggcggag gaggcgacgt
     2041 cgctagtgac atcgagaagc tgtgcaagaa ggcggaagag gaggccaagg aggcccagga
     2101 gaaggccgac gagctgagac aacgacaccc cgattcgcag gcggcggaag acgcggagga
     2161 tctcgcaaat ctggctgtcg ccgccgtgct gaccgcttgt ctgctggccc aggagcatcc
     2221 caacgccgat atcgctaagc tgtgcatcaa ggccgcgagc gaagctgcgg aggccgcgtc
     2281 aaaagccgcc gagctggccc aacggcaccc agacagtcaa gcagccagag atgctatcaa
     2341 actcgcctct caagccgccc gcgccgtgat cctggccatc atgctcgccg cagagaatcc
     2401 caacgctgat attgcaaagc tatgtattaa ggcggccagc gaagccgcag aggctgcctc
     2461 gaaagctgcg gaacttgcac agcggcaccc cgactcccag gccgcccgag atgcaattaa
     2521 actggccagt caggccgctg aggcggtgga gagagccatc tggctggccg ccgagaaccc
     2581 caatgccgat atcgccaaaa aatgcatcaa agcggcctcc gaagcagccg aggaggcctc
     2641 taaagctgcc gaagaggctc aaagacaccc agacagccag aaggcacgcg acgagatcaa
     2701 ggaggcctct cagaaggccg aggaggtgaa ggagagatgc aagagcgcca gcggaagtgg
     2761 cggaggaggc gacgtcgcta gcatgctcaa gatggagtct acacagcaga tggctagttc
     2821 gattattaac acttcttttg aagccgcagt cgttgctgcc acttcaactc ttgaacttat
     2881 gggcattcaa tatgattata atgaagttta tactagagtt aagagcaaat ttgattacgt
     2941 aatggatgat tcaggtgtta aaaacaatct gttgggtaaa gcggcaacta ttgatcaggc
     3001 actgaatgga aaatttggtt cagcagcaag aaatagaaat tggatggctg atactagaac
     3061 tacagctaga ctagatgaag atgtgaacaa acttagaatg atgctatcat ctaaaggaat
     3121 tgatcaaaaa atgagagtac ttaatgcatg tttcaatgtg aagcgagttc caggtaaatc
     3181 atcatcaata attaaatgta ctagattaat gagagataag attgaacgtg gtgaagtcga
     3241 agtggatgat tcgtttgttg aagaaaaaat ggaagttgac acaattgatt ggaaatctag
     3301 atatgaacaa ttggaaaaac gatttgaatc gctgaaacaa agggtgaatg aaaaatatac
     3361 aagttgggtg caaaaagcaa aaaaggtaaa tgaaaatatg tactctctcc aaaacgttat
     3421 atcacaacag cagagccaga ttgctgattt acaaaattac tgtaataaat tggaagttga
     3481 tctacaaaat aaaattagtt cacttgtgtc ttcagtcgaa tggtacctga aatcaatgga
     3541 attgcctgat gaaattaaga cagatattga acaacaacta aattccattg atgtaatcaa
     3601 tccaattaat gctatagatg attttgaatc actgatcaga aatattatac tcgactatga
     3661 cagaatattt ttaatgttta aaggattaat gaggcaatgc aactatgaat atacgtatga
     3721 ataggatccc ctgcagggct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3781 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3841 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3901 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3961 tgcggtgggc tctatggctt ctgaggataa gatacattga tgagtttgga caaaccacaa
     4021 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4081 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4141 aggttcaggg ggaggtgtgg gaggtttttt aaagcgcttg gtaccgtctt tccagtggtt
     4201 aattaaataa cttcgtatag catacattat acgaagttat ggatccgaac aaacgaccca
     4261 acacccgtgc gttttattct gtctttttat tgccgatcct cgaggccacc atgactagtg
     4321 cgtttaaact taagcttcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     4381 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4441 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4501 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4561 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4621 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4681 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4741 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4801 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4861 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4921 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4981 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5041 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     5101 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5161 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5221 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5281 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5341 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5401 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5461 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5521 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5581 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5641 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5701 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5761 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5821 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5881 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5941 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6001 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6061 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6121 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6181 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6241 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6301 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6361 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6421 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6481 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6541 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6601 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6661 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6721 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6781 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7441 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7681 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7741 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7801 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7861 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7921 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7981 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8041 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8101 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8161 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8221 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8281 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8341 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8401 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8461 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8521 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8581 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8641 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8701 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8761 aagtgccacc tgacgtc
//