LOCUS       sequence-7034-ex        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  AUTHORS   Colwill K, Wells CD, Elder K, Goudreault M, Hersi K, Kulkarni S, 
            Hardy WR, Pawson T, Morin GB
  TITLE     Modification of the Creator recombination system for proteomics 
            applications--improved expression by addition of splice sites.
  JOURNAL   BMC Biotechnol. 2006 Mar 6;6:13. doi: 10.1186/1472-6750-6-13.
  PUBMED    16519801
REFERENCE   2  (bases 1 to 4894)
  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: "BMC 
            Biotechnol."; date: "2006-03-6"; volume: "6:13. doi"; pages: " 
            10.1186/1472-6750-6-13"
COMMENT     Sequence Label: V1664 pLPS-3'RFP-mito - Acceptor
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    complement(597..630)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             843..1514
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVKMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERIMNFEDGGVV
                     TVTQDSTLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYKAKKQVQLPGAYKTDIKLDIISHNEDYTIVEQYERAEGRHST
                     GA"
     primer_bind     complement(970..988)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1682..1803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1719..1738)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1773..1792
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1810..2265)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1947..1968)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2159..2178
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2292..2396
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2398..2755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2398..2589
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2421..2441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2606..2741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2790..3584
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2790..3584
                     /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
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2844..2863)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3454..3473
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3772..3791)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3816..3863
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4192..4780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4681..4700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcataa
      601 cttcgtatag catacattat acgaagttat agatccaata ttattgaagc atttatcagg
      661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      721 ttccgcgcac atttccccga aaagtgccac ctgacgtgga tctcgagctc aagcttcgaa
      781 ttcagggttt ccttgacaat atcatactta tcctgtccct tttttttcca cagctaccgg
      841 tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt caaggtgaag atggagggct
      901 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      961 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     1021 tgtcccctca gttccagtac ggatccaagg cctacgtgaa gcaccccgcc gacatccccg
     1081 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcatcatg aacttcgagg
     1141 acggcggcgt ggtgaccgtg acccaggact ccaccctgca ggacggcgag ttcatctaca
     1201 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtgatgcag aagaagacca
     1261 tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga
     1321 tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct
     1381 acaaggccaa gaagcaggtc cagctgcctg gcgcctacaa gaccgacatc aagctggaca
     1441 tcatctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc
     1501 actccaccgg cgccctgtac aagaaactaa ttgctaaaag tgcagaagac gaaaaagcga
     1561 aggaagaacc agggaaccat acgacgttaa ttcttgcaat gttagctatt ggcgtgttct
     1621 ctttaggggc gtttatcaaa attattcaat taagaaaaaa taattaacaa ttgttgttgt
     1681 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1741 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1801 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1861 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1921 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1981 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2041 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2101 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2161 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2221 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2341 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2401 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2521 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2581 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2641 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2701 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     2761 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2821 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2881 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2941 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3001 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3061 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3121 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3181 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3241 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3301 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3361 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3421 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3481 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3541 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3601 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3661 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3721 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     3781 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3841 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3901 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3961 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4021 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4321 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4801 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4861 tgattctgtg gataaccgta ttaccgccat gcat
//
