LOCUS       sequence-7354-ex        6161 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 6161)
  AUTHORS   Trivedi RN, Almeida KH, Fornsaglio JL, Schamus S, Sobol RW
  TITLE     The role of base excision repair in the sensitivity and resistance 
            to temozolomide-mediated cell death.
  JOURNAL   Cancer Res. 2005 Jul 15. 65(14):6394-400.
  PUBMED    16024643
REFERENCE   2  (bases 1 to 6161)
  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: "Cancer Res.
            2005 Jul 15. 65(14):6394-400."
COMMENT     Sequence Label: pRS1422
FEATURES             Location/Qualifiers
     source          1..6161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1917..2146
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2129..2148
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2204..2778,2779..2781,2782..2790)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2371..2388)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2598..2617
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2813..3616
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2876..2895)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3486..3505
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3648..3665)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3654..3878
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3925..3944
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4004..4020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4004..4020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4017..4039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4028..4044
                     /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(4052..4058,4059..4076,4077..4082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4097..4118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4235..4252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4406..4994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4486..4505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5165..5956,5957..6025))
                     /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     5788..5807
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6026..6130)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatgatcca tggtcacccc cgctttgcag atgaagaaac caaagcagtt
      961 ttgccgacgg atggggcaaa agaagcagcg accagctaga gcagggcagc cacacagctc
     1021 gtccgacgca gcccaggcac ctgcagagca gccacacagc tcgtccgatg cagcccaggc
     1081 accttgcccc agggagcgct gcttgggacc gcccaccact ccgggcccat accgcagcat
     1141 ctatttctca agcccaaagg gccaccttac ccgactgggg ttggagttct tcgaccagcc
     1201 ggcagtcccc ctggcccggg catttctggg acaggtccta gtccggcgac ttcctaatgg
     1261 cacagaactc cgaggccgca tcgtggagac cgaggcatac ctggggccag aggatgaagc
     1321 cgcccactca aggggtggcc ggcagacccc ccgcaaccga ggcatgttca tgaagccggg
     1381 gaccctgtac gtgtacatca tttacggcat gtacttctgc atgaacatct ccagccaggg
     1441 ggacggggct tgcgtcttgc tgcgagcact ggagcccctg gaaggtctgg agaccatgcg
     1501 tcagcttcgc agcaccctcc ggaaaggcac cgccagccgt gtcctcaagg accgcgagct
     1561 ctgcagtggc ccctccaagc tgtgccaggc cctggccatc aacaagagct ttgaccagag
     1621 ggacctggca caggatgaag ctgtatggct ggagcgtggt cccctggagc ccagtgagcc
     1681 ggctgtagtg gcagcagccc gggtgggcgt cggccatgca ggggagtggg cccggaaacc
     1741 cctccgcttc tatgtccggg gcagcccctg ggtcagtgtg gtcgacagag tggctgagca
     1801 ggacacacag gcctgactcg aatctgcggc cgcgtcgacg gaattcagtg gatccactag
     1861 taacggccgc cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga
     1921 gtactccctc tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga
     1981 ggaggatttg atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg
     2041 gtcagaaaag acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc
     2101 atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag
     2161 gtccaactgc aggtcgagca tgcatctagg gcggccaatt ccgcccctct ccctcccccc
     2221 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2281 gattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2341 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     2401 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2461 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2521 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2581 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2641 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2701 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2761 gaaaaacacg atgataagct tgccacaacc cgggataatt cctgcagcca atatgggatc
     2821 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2881 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2941 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3001 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3061 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3121 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3181 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3241 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3301 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3361 cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3421 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3481 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3541 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3601 tgacgagttc ttctgagggg atcaattctc tagagctcgc tgatcagcct cgactgtgcc
     3661 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3721 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3781 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3841 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3901 ctggggctcg agtgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3961 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4021 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4081 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4201 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4381 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     4621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5101 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5161 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5221 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5281 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5341 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5401 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5461 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5521 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5581 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5641 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5701 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5761 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5821 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5881 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5941 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6001 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6061 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6121 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
