pLZ299

LOCUS       pLZ299         7837 bp DNA     circular SYN 10-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7837)
  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..7837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     166..191
                     /label=182-WKG-596-F
     misc_feature    232
                     /label=CMV
     promoter        245..441
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        470..474
                     /label=P_TAL
     misc_recomb     490..523
                     /label=LoxP
     protein_bind    complement(490..523)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    579..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      812..996
                     /label=SV40 late polyA
     polyA_signal    814..935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1033..1066
                     /label=loxP
     protein_bind    complement(1033..1066)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1073..1121
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      1141..1150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1147..1209
                     /label=Igk secretion signal
     CDS             1147
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /translation=""
     CDS             1150
                     /codon_start=1
                     /label=mCherry
                     /translation=""
     CDS             1216..1803
                     /codon_start=1
                     /label=NS3a H1
                     /translation="KKKGSVVIVGRINLSGDTAYSQQTRGLEGCQETSQTGRDKNQVEG
                     EVQVVSTATQSFLATSINGVLWTVYHGAGTRTIASPKGPVTQMYTNVDKDLVGWQAPQG
                     SRSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGH
                     AVGIFRAAVSTRGVAKAVDFIPVESLETTMRSP"
     misc_feature    1810..2553
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     primer_bind     complement(1877..1899)
                     /label=186-WKG-603-R
     misc_feature    2083..2085
                     /label=F93A
                     /note="F93A"
     misc_feature    2537..2553
                     /label=EpoR-D1_D2_TM
     misc_feature    2563..3391
                     /label=IL6ST
     misc_feature    2590..2616
                     /label=box1
     misc_feature    2914..2916
                     /label=Y759A
     polyA_signal    3434..3658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3704..4132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3742)
                     /label=184-WKG-599-R
     promoter        4146..4475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4542..5336
                     /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"
     polyA_signal    5510..5631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5676..5696)
                     /label=M13-rev
     primer_bind     complement(5680..5696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(5702..5724)
                     /label=LacO
     protein_bind    5704..5720
                     /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(5728..5758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(5729..5758)
                     /label=lac
     protein_bind    5773..5794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    5900..6840
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(6841..7701)
                     /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(7702..7806)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(7743..7771)
                     /label=Amp prom
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 cgtgatctgc
      241 gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
      301 ctaactccgc ccagttccgc ccattctccg ccccatcgct gactaatttt ttttatttat
      361 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
      421 ggaggcctag gcttttgcaa aaagctacga atcgcgctag tggccggccg aagcagaagg
      481 ctagcgctta taacttcgta tagcatacat tatacgaagt tatcggcgcg ccaatgctcc
      541 gaatgcatca gtgcccctgc agggctgatc agcctcgact gtgccttcta gttgccagcc
      601 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
      661 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
      721 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
      781 tggggatgcg gtgggctcta tggcttctga ggataagata cattgatgag tttggacaaa
      841 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      901 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      961 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag cgcttggtac cgtctttcca
     1021 gtggttaatt aaataacttc gtatagcata cattatacga agttatcgat ccgaacaaac
     1081 gacccaacac ccgtgcgttt tattctgtct ttttattgcc gatcctcgag gaattcgccg
     1141 gccaccatgg agaccgatac cttattactg tgggtgcttc ttctgtgggt tcccggtagc
     1201 accggagacg gatccaagaa gaagggcagc gtggtgatcg tgggcagaat caacctgagc
     1261 ggcgacaccg cctacagcca gcagaccaga ggcctggagg gctgccagga gaccagccag
     1321 accggcagag acaagaacca ggtggagggc gaggtgcagg tggtgagcac cgccacccag
     1381 agcttcctgg ccaccagcat caacggcgtg ctgtggaccg tgtaccacgg cgccggcacc
     1441 agaaccatcg ccagccccaa gggccccgtg acccagatgt acacgaacgt ggacaaggac
     1501 ttagtgggct ggcaggcccc ccaaggcagc agaagcctga ccccctgcac ctgcggcagc
     1561 agcgacctgt acctggtgac cagacacgcc gacgtgatcc ccgtgagaag aagaggcgac
     1621 agcagaggca gcctgctgag ccccagaccc atcagctatc tgaaaggctc aagcggcggc
     1681 cccctgttgt gtcccgccgg ccatgccgtg ggcatcttca gagccgccgt gagcaccaga
     1741 ggcgtggcca aggccgtgga cttcatcccc gtggagagcc tggagaccac catgagaagc
     1801 cccgaattcg caccttcacc cagcctcccg gaccccaagt ttgagagcaa agcggccctg
     1861 ctggcatccc ggggctccga agaacttctg tgcttcaccc aacgcttgga agacttggtg
     1921 tgtttctggg aggaagcggc gagctccggg atggacttca actacagctt ctcataccag
     1981 ctcgagggtg agtcacgaaa gtcatgtagc ctgcaccagg ctcccaccgt ccgcggctcc
     2041 gtgcgtttct ggtgttcact gccaacagcg gacacatcga gtgctgtgcc gctggagctg
     2101 caggtgacgg aggcgtccgg ttctcctcgc tatcaccgca tcatccatat caatgaagta
     2161 gtgctcctgg acgcccccgc ggggctgctg gcgcgccggg cagaagaggg cagccacgtg
     2221 gtgctgcgct ggctgccacc tcctggagca cctatgacca cccacatccg atatgaagtg
     2281 gacgtgtcgg caggcaaccg ggcaggaggg acacaaaggg tggaggtcct ggaaggccgc
     2341 actgagtgtg ttctgagcaa cctgcggggc gggacgcgct acaccttcgc tgttcgagcg
     2401 cgcatggccg agccgagctt cagcggattc tggagtgcct ggtctgagcc cgcgtcacta
     2461 ctgaccgcta gcgacctgga ccctctcatc ttgacgctgt ctctcattct ggtcctcatc
     2521 tcgttgttgc tgacggttct ggccctgctg tccgcagcag caaataagcg agacctaatt
     2581 aaaaaacaca tctggcctaa tgttccagat ccttcaaaga gtcatattgc ccagtggtca
     2641 cctcacactc ctccaaggca caattttaat tcaaaagatc aaatgtattc agatggcaat
     2701 ttcactgatg taagtgttgt ggaaatagaa gcaaatgaca aaaagccttt tccagaagat
     2761 ctgaaatcat tggacctgtt caaaaaggaa aaaattaata ctgaaggaca cagcagtggt
     2821 attggggggt cttcatgcat gtcatcttct aggccaagca tttctagcag tgatgaaaat
     2881 gaatcttcac aaaacacttc gagcactgtc caggcttcta ccgtggtaca cagtggctac
     2941 agacaccaag ttccgtcagt ccaagtcttc tcaagatccg agtctaccca gcccttgtta
     3001 gattcagagg agcggccaga agatctacaa ttagtagatc atgtagatgg cggtgatggt
     3061 atttttccca ggcaacagta cttcaaacag aactgcagtc agcatgaatc cagtccagat
     3121 atttcacatt ttgaaaggtc aaagcaagtt tcatcagtca atgaggaaga ttttgttaga
     3181 cttaaacagc agatttcaga tcatatttca caatcctgtg gatctgggca aatgaaaatg
     3241 tttcaggaag tttctgcagc agatgctttt ggtccaggta ctgagggaca agtagaaaga
     3301 tttgaaacag ttggcatgga ggctgcgact gatgaaggca tgcctaaaag ttacttacca
     3361 cagactgtac ggcaaggcgg ctacatgcct cagtaatcta gagggcccgt ttaaacccgc
     3421 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3481 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3541 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3601 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3661 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3721 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3781 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3841 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3901 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3961 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4021 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4081 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4141 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4201 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4261 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4321 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4381 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4441 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4501 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4561 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4621 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4681 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4741 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4801 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4861 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4921 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4981 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5041 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5101 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5161 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5221 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5281 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5341 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5401 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5461 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5521 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5581 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5641 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5701 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5761 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5821 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5881 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6001 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6241 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6481 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6661 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6721 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6781 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6841 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6901 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6961 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7021 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7081 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7141 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7201 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7261 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7321 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7381 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7441 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7501 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7561 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7621 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7681 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7741 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7801 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//