pLZ362

LOCUS       pLZ362               6169 bp DNA     circular SYN 20-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6169)
  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..6169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..232
                     /label=oSCL771
     misc_feature    232
                     /label=CMV
     promoter        245..441
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     primer_bind     complement(425..461)
                     /label=oSCL1562
     primer_bind     444..489
                     /label=oLH099 LoxP F
     primer_bind     444..469
                     /label=oSCL1563
     promoter        470..474
                     /label=P_TAL
     primer_bind     479..524
                     /label=olyl285
     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)."
     primer_bind     527..561
                     /label=olyl405
     primer_bind     complement(531..557)
                     /label=olyl406
     primer_bind     564..583
                     /label=olyl294
     polyA_signal    579..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(790..811)
                     /label=olyl286
     primer_bind     792..844
                     /label=olyl287
     terminator      812..996
                     /label=SV40 late polyA
     primer_bind     complement(977..1001)
                     /label=olyl288
     primer_bind     982..1023
                     /label=olyl289
     primer_bind     1033..1067
                     /label=olyl285
     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)"
     primer_bind     complement(1093..1124)
                     /label=olyl290
     primer_bind     complement(1115..1157)
                     /label=oLH100 LoxP R
     primer_bind     1137..1161
                     /label=oSCL1563
     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=""
     primer_bind     1167..1186
                     /label=oYS520
     primer_bind     complement(1189..1215)
                     /label=oSCL1564
     primer_bind     1196..1240
                     /label=oLH200 Nluc F
     misc_feature    1219..1728
                     /label=NLuc (nano Luciferase)
                     /label=nonstandard type:  reporter
     misc_feature    1249..1251
                     /label=cyt-STOP (TGG->TGA)
     primer_bind     complement(1711..1751)
                     /label=oLH201 Nluc R
     primer_bind     complement(1711..1728)
                     /label=oYS12
     primer_bind     complement(1733..1754)
                     /label=oLeo540_3.1_SfiI_R
     primer_bind     complement(1740..1757)
                     /label=oYW220
                     /note="RA-(XbaI-Coh2) PCR-R"
     primer_bind     1754..1773
                     /label=olyl294
     polyA_signal    1769..1993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1980..2001)
                     /label=olyl286
     primer_bind     1982..2002
                     /label=olyl287
     rep_origin      2039..2467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2481..2810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2794..2815)
                     /label=oSCL1562
     CDS             2877..3671
                     /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    3845..3966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3936..3968)
                     /label=olyl287
     primer_bind     complement(4011..4031)
                     /label=M13-rev
     primer_bind     complement(4015..4031)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(4037..4059)
                     /label=LacO
     protein_bind    4039..4055
                     /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(4063..4093)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(4064..4093)
                     /label=lac
     protein_bind    4108..4129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    4235..5172
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(5173..6033)
                     /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(6034..6138)
                     /gene="bla"
                     /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 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 gatccatggt cttcacactc gaagatttcg ttggggactg gcgacagaca
     1261 gccggctaca acctggacca agtccttgaa cagggaggtg tgtccagttt gtttcagaat
     1321 ctcggggtgt ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa tgggctgaag
     1381 atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc
     1441 gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgcac
     1501 tatggcacac tggtaatcga cggggttacg ccgaacatga tcgactattt cggacggccg
     1561 tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac
     1621 ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta
     1681 accatcaacg gagtgaccgg ctggcggctg tgcgaacgca ttctggcgta atctagaggg
     1741 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1801 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1861 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1921 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1981 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2101 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2221 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2341 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2401 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2461 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2521 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2581 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2641 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2701 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2761 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2821 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2881 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2941 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3001 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3061 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3121 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3181 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3241 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3301 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3361 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3421 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3481 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3541 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3601 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3661 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3721 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3781 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3841 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3901 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3961 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4021 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4081 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4141 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4201 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4261 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4921 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     4981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5461 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5761 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//