LOCUS       sequence-229780-        6383 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 6383)
  AUTHORS   Bhatt A, Kaverina I, Otey C, Huttenlocher A
  TITLE     Regulation of focal complex composition and disassembly by the 
            calcium-dependent protease calpain.
  JOURNAL   J Cell Sci. 2002 Sep 1. 115(Pt 17):3415-25.
  PUBMED    12154072
REFERENCE   2  (bases 1 to 6383)
  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: "J Cell Sci.
            2002 Sep 1. 115(Pt 17):3415-25."
COMMENT     Sequence Label: RFP-zyxin
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2418..2427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2424..2426,2427..2429,2430..3104)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(2607..2627)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2825..2844
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3019..3042
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3225..3346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3262..3281)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3316..3335
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3353..3808)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3490..3511)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3702..3721
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3835..3939
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3941..4298
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3941..4132
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3964..3984)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4149..4284
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4333..5127
                     /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             4333..5127
                     /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(4387..4406)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4997..5016
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5315..5334)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5359..5406
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5735..6323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6224..6243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      661 tcagatctcg agctcaagct tcgaattcca tggcggcccc ccgcccgtct cccgcgatct
      721 ccgtttcggt ctcggctccg gctttttacg ccccgcagaa gaagttcggc cctgtggtgg
      781 ccccaaagcc caaagtgaat cccttccggc ccggggacag cgagcctccc ccggcacccg
      841 gggcccagcg cgcacagatg ggccgggtgg gcgagattcc cccgccgccc ccggaagact
      901 ttcccctgcc tccacctccc cttgctgggg atggcgacga tgcagagggt gctctgggag
      961 gtgccttccc gccgccccct cccccgatcg aggaatcatt tccccctgcg cctctggagg
     1021 aggagatctt cccttccccg ccgcctcctc cggaggagga gggagggcct gaggccccca
     1081 taccgccccc accacagccc agggagaagg tgagcagtat tgatttggag atcgactctc
     1141 tgtcctcact gctggatgac atgaccaaga atgatccttt caaagcccgg gtgtcatctg
     1201 gatatgtgcc cccaccagtg gccactccat tcagttccaa gtccagtacc aagcctgcag
     1261 ccgggggcac agcacccctg cctccttgga agtccccttc cagctcccag cctctgcccc
     1321 aggttccggc tccggctcag agccagacac agttccatgt tcagccccag ccccagccca
     1381 agcctcaggt ccaactccat gtccagtccc agacccagcc tgtgtctttg gctaacaccc
     1441 agccccgagg gcccccagcc tcatctccgg ctccagcccc taagttttct ccagtgactc
     1501 ctaagtttac tcctgtggct tccaagttca gtcctggagc cccaggtgga tctgggtcac
     1561 aaccaaatca aaaattgggg caccccgaag ctctttctgc tggcacaggc tcccctcaac
     1621 ctcccagctt cacctatgcc cagcagaggg agaagccccg agtgcaggag aagcagcacc
     1681 ccgtgccccc accggctcag aaccaaaacc aggtgcgctc ccctggggcc ccagggcccc
     1741 tgactctgaa ggaggtggag gagctggagc agctgaccca gcagctaatg caggacatgg
     1801 agcatcctca gaggcagaat gtggctgtca acgaactctg cggccgatgc catcaacccc
     1861 tggcccgggc gcagccagcc gtccgcgctc tagggcagct gttccacatc gcctgcttca
     1921 cctgccacca gtgtgcgcag cagctccagg gccagcagtt ctacagtctg gagggggcgc
     1981 cgtactgcga gggctgttac actgacaccc tggagaagtg taacacctgc ggggagccca
     2041 tcactgaccg catgctgagg gccacgggca aggcctatca cccgcactgc ttcacctgtg
     2101 tggtctgcgc ccgccccctg gagggcacct ccttcatcgt ggaccaggcc aaccggcccc
     2161 actgtgtccc cgactaccac aagcagtacg ccccgaggtg ctccgtctgc tctgagccca
     2221 tcatgcctga gcctggccga gatgagactg tgcgagtggt cgccctggac aagaacttcc
     2281 acatgaagtg ttacaagtgt gaggactgcg ggaagcccct gtcgattgag gcagatgaca
     2341 atggctgctt ccccctggac ggtcacgtgc tctgtcggaa gtgccacact gctagagccc
     2401 agacggatcc accggtcgcc accatggtgc gctcctccaa gaacgtcatc aaggagttca
     2461 tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg
     2521 agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg
     2581 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
     2641 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
     2701 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     2761 ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg
     2821 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     2881 cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc
     2941 actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct
     3001 actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     3061 agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg cgactctaga
     3121 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3181 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3241 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3301 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3361 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3421 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3481 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3541 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3601 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3661 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3721 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3781 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3961 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4141 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4201 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4261 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4321 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4381 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4441 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4501 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4561 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4621 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4681 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4741 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4801 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4861 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4921 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4981 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5041 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5101 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5161 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5221 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5281 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5341 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5401 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5461 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5521 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5581 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5641 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5701 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5881 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6361 tttgctggcc ttttgctcac atg
//
