LOCUS       sequence-469991-        6276 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 6276)
  AUTHORS   Soutoglou E, Dorn JF, Sengupta K, Jasin M, Nussenzweig A, Ried T, 
            Danuser G, Misteli T
  TITLE     Positional stability of single double-strand breaks in mammalian 
            cells.
  JOURNAL   Nat Cell Biol. 2007 Jun . 9(6):675-82.
  PUBMED    17486118
REFERENCE   2  (bases 1 to 6276)
  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: "Nat Cell 
            Biol. 2007 Jun . 9(6):675-82."
COMMENT     Sequence Label: ISceI-GR-RFP
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1294
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     primer_bind     1018..1037
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3071..3192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3108..3127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3162..3181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3199..3654)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3336..3357)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3548..3567
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3681..3785
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3787..4144
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3787..3978
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3810..3830)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3995..4130
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4179..4973
                     /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             4179..4973
                     /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(4233..4252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4843..4862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5161..5180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5205..5252
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5581..6169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6070..6089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggacaacac cgaggacgtc atcaaggagt tcatgcagtt
      661 caaggtgcgc atggagggct ccgtgaacgg ccactacttc gagatcgagg gcgagggcga
      721 gggcaagccc tacgagggca cccagaccgc caagctgcag gtgaccaagg gcggccccct
      781 gcccttcgcc tgggacatcc tgtcccccca gttccagtac ggctccaagg cctacgtgaa
      841 gcaccccgcc gacatccccg actacatgaa gctgtccttc cccgagggct tcacctggga
      901 gcgctccatg aacttcgagg acggcggcgt ggtggaggtg cagcaggact cctccctgca
      961 ggacggcacc ttcatctaca aggtgaagtt caagggcgtg aacttccccg ccgacggccc
     1021 cgtaatgcag aagaagactg ccggctggga gccctccacc gagaagctgt acccccagga
     1081 cggcgtgctg aagggcgaga tctcccacgc cctgaagctg aaggacggcg gccactacac
     1141 ctgcgacttc aagaccgtgt acaaggccaa gaagcccgtg cagctgcccg gcaaccacta
     1201 cgtggactcc aagctggaca tcaccaacca caacgaggac tacaccgtgg tggagcagta
     1261 cgagcacgcc gaggcccgcc actccggctc ccagtccgga ctcagatctc gagctcaaat
     1321 gaaaaacatc aaaaaaaacc aggtaatgaa cctgggtccg aactctaaac tgctgaaaga
     1381 atacaaatcc cagctgatcg aactgaacat cgaacagttc gaagcaggta tcggtctgat
     1441 cctgggtgat gcttacatcc gttctcgtga tgaaggtaaa acctactgta tgcagttcga
     1501 gtggaaaaac aaagcataca tggaccacgt atgtctgctg tacgatcagt gggtactgtc
     1561 cccgccgcac aaaaaagaac gtgttaacca cctgggtaac ctggtaatca cctggggcgc
     1621 ccagactttc aaacaccaag ctttcaacaa actggctaac ctgttcatcg ttaacaacaa
     1681 aaaaaccatc ccgaacaacc tggttgaaaa ctacctgacc ccgatgtctc tggcatactg
     1741 gttcatggat gatggtggta aatgggatta caacaaaaac tctaccaaca aatcgatcgt
     1801 actgaacacc cagtctttca ctttcgaaga agtagaatac ctggttaagg gtctgcgtaa
     1861 caaattccaa ctgaactgtt acgtaaaaat caacaaaaac aaaccgatca tctacatcga
     1921 ttctatgtct tacctgatct tctacaacct gatcaaaccg tacctgatcc cgcagatgat
     1981 gtacaaactg ccgaacacta tctcctccga aactttcctg aaatgtcgac ggtaccgcgg
     2041 gtatcggaaa tgtcttcagg ctggaatgaa ccttgaagct cgaaaaacaa agaaaaaaat
     2101 caaagggatt cagcaagcca ctgcaggagt ctcacaagac acttcggaaa atcctaacaa
     2161 aacaatagtt cctgcagcat taccacagct cacccctacc ttggtgtcac tgctggaggt
     2221 gattgaaccc gaggtgttgt atgcaggata tgatagctct gttccagatt cagcatggag
     2281 aattatgacc acactcaaca tgttaggtgg gcgtcaagtg attgcagcag tgaaatgggc
     2341 aaaggcgata ccaggcttca gaaacttaca cctggatgac caaatgaccc tgctacagta
     2401 ctcatggatg tttctcatgg catttgccct gggttggaga tcatacagac aatcaagtgg
     2461 aaacctgctc tgctttgctc ctgatctgat tattaatgag cagagaatgt ctctacccgg
     2521 catgtatgac caatgtaaac acatgctgtt tgtctcctct gaattacaaa gattgcaggt
     2581 atcctatgaa gagtatctct gtatgaaaac cttactgctt ctctcctcag ttcctaagga
     2641 aggtctgaag agccaagagt tatttgatga gattcgaatg acttatatca aagagctagg
     2701 aaaagccatc gtcaaaaggg aagggaactc cagtcagaac tggcaacggt tttaccaact
     2761 gacaaagctt ctggactcca tgcatgaggt ggttgagaat ctccttacct actgcttcca
     2821 gacatttttg gataagacca tgagtattga attcccagag atgttagctg aaatcatcac
     2881 taatcagata ccaaaatatt caaatggaaa tatcaaaaag cttctgtttc atcaaaaatt
     2941 tggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     3001 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3061 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3181 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3241 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3301 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3361 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3421 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3481 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3541 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3601 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3781 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3901 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3961 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4021 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4081 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4141 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4201 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4261 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4321 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4381 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4441 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4501 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4561 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4621 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4681 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4741 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4801 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4981 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5041 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5101 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5161 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5221 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5281 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5341 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5401 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5701 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6241 gttatcccct gattctgtgg ataaccgtat taccgc
//
