LOCUS       sequence-7033-ex        4840 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 4840)
  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 4840)
  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: V1663 pLPS-3'RFP - Acceptor
FEATURES             Location/Qualifiers
     source          1..4840
                     /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..1517
                     /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"
     primer_bind     1238..1257
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1628..1749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1665..1684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1719..1738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1756..2211)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1893..1914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2105..2124
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2238..2342
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2344..2701
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2344..2535
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2367..2387)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2552..2687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2736..3530
                     /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             2736..3530
                     /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(2790..2809)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3400..3419
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3718..3737)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3762..3809
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4138..4726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4627..4646
                     /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 cgtaatgcag 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 cgcctaatct agatcataat cagccatacc acatttgtag aggttttact
     1561 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1621 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1741 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1801 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     1861 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     1921 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     1981 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2041 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2101 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2161 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2221 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2281 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2341 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2401 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2461 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2521 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2581 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2641 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2701 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2761 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2821 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2881 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     2941 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3001 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3061 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3121 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3181 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3241 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3301 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3361 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3421 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3481 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3541 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3601 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3661 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     3721 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3781 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3841 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3901 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3961 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4021 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4081 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4261 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4801 atcccctgat tctgtggata accgtattac cgccatgcat
//
