LOCUS       sequence-59603-e        6343 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 6343)
  AUTHORS   Gunn A, Bennardo N, Cheng A, Stark JM
  TITLE     Correct end use during end joining of multiple chromosomal double 
            strand breaks is influenced by repair protein RAD50, DNA-dependent 
            protein kinase DNA-PKcs, and transcription context.
  JOURNAL   J Biol Chem. 2011 Dec 9;286(49):42470-82. doi: 
            10.1074/jbc.M111.309252. Epub 2011 Oct 24.
  PUBMED    22027841
REFERENCE   2  (bases 1 to 6343)
  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"; doi: 
            "10.1074/jbc.M111.309252"; journalName: "J Biol Chem"; date: 
            "2011-12-9- 9"; volume: "286"; issue: "49"; pages: "42470-82"
COMMENT     Sequence Label: pCAGGS-I-SceI-Trex2
FEATURES             Location/Qualifiers
     source          1..6343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1765..1785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1807..1827
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1846..1872
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3355..3374)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3420..3475
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3474..3493)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3846..3862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3846..3862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3859..3881)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3870..3886
                     /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(3894..3900,3901..3918,3919..3924))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3939..3960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4019..4214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4065..4200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4220..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4257..4276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4311..4330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4422..4439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4593..5181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4673..4692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5352..6143,6144..6212))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5975..5994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6213..6317)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgaa attcagatcc
     1741 gccgccacta tgggatcaag atcgccaaaa aagaagagaa aggtgccgaa gaagcatgca
     1801 gcaccaccaa aaaaaaaacg aaaagtagaa gacccacgat ttatgtaccc atacgatgtt
     1861 cctgactatg cgggtatgaa aaacatcaaa aaaaaccagg taatgaacct gggtccgaac
     1921 tctaaactgc tgaaagaata caaatcccag ctgatcgaac tgaacatcga acagttcgaa
     1981 gcaggtatcg gtctgatcct gggtgatgct tacatccgtt ctcgtgatga aggtaaaacc
     2041 tactgtatgc agttcgagtg gaaaaacaaa gcatacatgg accacgtatg tctgctgtac
     2101 gatcagtggg tactgtcccc gccgcacaaa aaagaacgtg ttaaccacct gggtaacctg
     2161 gtaatcacct ggggcgccca gactttcaaa caccaagctt tcaacaaact ggctaacctg
     2221 ttcatcgtta acaacaaaaa aaccatcccg aacaacctgg ttgaaaacta cctgaccccg
     2281 atgtctctgg catactggtt catggatgat ggtggtaaat gggattacaa caaaaactct
     2341 accaacaaat cgatcgtact gaacacccag tctttcactt tcgaagaagt agaatacctg
     2401 gttaagggtc tgcgtaacaa attccaactg aactgttacg taaaaatcaa caaaaacaaa
     2461 ccgatcatct acatcgattc tatgtcttac ctgatcttct acaacctgat caaaccgtac
     2521 ctgatcccgc agatgatgta caaactgccg aacactatct cctccgaaac tttcctgaaa
     2581 agatcttgtt cctgtgagaa catcagcatg tctgagccac cccgggctga gacctttgta
     2641 ttcctggacc tagaagccac tgggctccca aacatggatc ctgagattgc agagatatcc
     2701 ctttttgctg ttcaccgctc ttccctggag aacccagaac gggatgattc tggttccttg
     2761 gtgctgcccc gtgttctgga caagctcaca ctgtgcatgt gcccggagcg cccctttact
     2821 gccaaggcca gtgagattac tggtttgagc agcgaaagcc tgatgcactg cgggaaggct
     2881 ggtttcaatg gcgctgtggt aaggacactg cagggcttcc taagccgcca ggagggcccc
     2941 atctgccttg tggcccacaa tggcttcgat tatgacttcc cactgctgtg cacggagcta
     3001 caacgtctgg gtgcccatct gccccaagac actgtctgcc tggacacact gcctgcattg
     3061 cggggcctgg accgtgctca cagccacggc accagggctc aaggccgcaa aagctacagc
     3121 ctggccagtc tcttccaccg ctacttccag gctgaaccca gtgctgccca ttcagcagaa
     3181 ggtgatgtgc acaccctgct tctgatcttc ctgcatcgtg ctcctgagct gctcgcctgg
     3241 gcagatgagc aggcccgcag ctgggctcat attgagccca tgtacgtgcc acctgatggt
     3301 ccaagcctcg aagcctgaat gccagtgcca tccattcatc caggctctcg aggatctttt
     3361 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3421 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3481 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3541 ggcaacatat gccatatgct ggctgccatg aacaaaggtg gctataaaga ggtcatcagt
     3601 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3661 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3721 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3781 ctcttctctt atgaagatcc ctcgacctgc agcccaagct gcggccgcag cttggcgtaa
     3841 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3901 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3961 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     4021 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4081 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4141 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4201 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4261 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4321 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     4381 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaaag ctcacgctgt
     4801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5401 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5461 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5521 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5581 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5641 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5701 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5761 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5821 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5881 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5941 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6001 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6061 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6121 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6181 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6241 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6301 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
