LOCUS       sequence-454076-        4617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0, N-terminal tag module (CCAT-AATG) containing 
            LoxP-mCherry-stop..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4617)
  AUTHORS   Chiang BJ, Lin KY, Chen YF, Huang CY, Goh FJ, Huang LT, Chen LH, Wu 
            CH
  TITLE     Development of a tightly regulated copper-inducible transient gene 
            expression system in Nicotiana benthamiana incorporating suicide 
            exon and Cre recombinase
  JOURNAL   bioRxiv 2024.03.23.586378
REFERENCE   2  (bases 1 to 4617)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.23.586378"
COMMENT     Sequence Label: pCHW110035
FEATURES             Location/Qualifiers
     source          1..4617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(260..278)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    887..1061
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(1071..1104)
                     /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(1184..1200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1184..1200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1197..1219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1208..1224
                     /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(1232..1238,1239..1256,1257..1262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1277..1298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1415..1432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1586..2173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1666..1685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2502..2549
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2574..2593
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2781..3575)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLCGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2892..2911)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3502..3521
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3610..3967)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3624..3759
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3776..3967
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3924..3944
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3969..4073)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4141..4159
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4197..4219)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4319..4338
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4532..4554
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(join(4617,1..33))
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 taacttcgta tagcatacat tatacgaagt tatattaagg gttgaatgaa aactaatctt
       61 tttctctttc tcatcttttc acttctccta tcattatcct cggccgaatt cggaggtgtg
      121 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
      181 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      241 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
      301 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      361 gacatccccg actacttgaa gctgtccttc cccgagtaat agtgaggctt caagtgggag
      421 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      481 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
      541 gtaatgcaga agaaaaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
      601 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
      661 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
      721 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
      781 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gggtggccat
      841 gatgagcttt aagcttctct agctagagtc gatcgacaag ctcgagtttc tccataataa
      901 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
      961 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
     1021 tctaattcct aaaaccaaaa tccagtacta aaatccagat cgctgaccta ataacttcgt
     1081 atagcataca ttatacgaag ttatattaag ggttgaatgt gagaccatcg gatcccgggc
     1141 ccgtcgactg cagaggcctg catgcaagct tggtgtaatc atggtcatag ctgtttcctg
     1201 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1261 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1321 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1381 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1681 ttcccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2281 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     2341 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     2401 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     2461 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     2521 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     2581 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     2641 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     2701 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     2761 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     2821 tgcgaatcgg gagcggcgat accgcaaagc acgaggaagc ggtcagccca ttcgccgcca
     2881 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     2941 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     3001 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     3061 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     3121 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     3181 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     3241 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     3301 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     3361 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     3421 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     3481 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     3541 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     3601 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     3661 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     3721 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     3781 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     3841 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     3901 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     3961 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4081 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4141 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4201 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4261 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     4321 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4381 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     4441 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     4501 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     4561 agagaattcg agctcggtac ctcgcgaata catctagatg gtctctccat gacctaa
//
