LOCUS       sequence-214603-        6439 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 6439)
  AUTHORS   Unsal-Kacmaz K, Chastain PD, Qu PP, Minoo P, Cordeiro-Stone M, 
            Sancar A, Kaufmann WK
  TITLE     The human Tim/Tipin complex coordinates an Intra-S checkpoint 
            response to UV that slows replication fork displacement.
  JOURNAL   Mol Cell Biol. 2007 Apr . 27(8):3131-42.
  PUBMED    17296725
REFERENCE   2  (bases 1 to 6439)
  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: "Mol Cell 
            Biol. 2007 Apr . 27(8):3131-42."
COMMENT     Sequence Label: pcDNA4-Chk1-Flag
FEATURES             Location/Qualifiers
     source          1..6439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        85..414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      265..400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     327..346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        462..509
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             528..902
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1032..1153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1069..1088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1123..1142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1202..1218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1202..1218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1215..1237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1226..1242
                     /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(1250..1256,1257..1274,1275..1280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1295..1316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1433..1450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1604..2192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1684..1703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2363..3154,3155..3223))
                     /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     2986..3005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3224..3328)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3403..3422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3594..3973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3974..4177
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4128..4148
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4222..4241
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4222..4240
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5754..5777
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5806..5823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5812..6036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6082..6439,1..71)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6169..6188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6379..6400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
       61 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
      181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      241 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      301 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
      361 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
      421 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
      481 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
      541 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
      601 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
      661 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
      721 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
      781 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
      841 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
      901 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
      961 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     1021 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1081 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1141 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     1201 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1261 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1321 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1381 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1441 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1501 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1561 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1621 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1681 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1741 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1801 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1861 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1921 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1981 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2041 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2101 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2161 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2221 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3301 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     3361 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     3421 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     3481 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     3541 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     3601 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     3661 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3721 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3781 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3841 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3901 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3961 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     4021 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     4081 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     4141 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
     4201 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagtt
     4261 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcctc gagatggcag
     4321 tgccctttgt ggaagactgg gacttggtgc aaaccctggg agaaggtgcc tatggagaag
     4381 ttcaacttgc tgtgaataga gtaactgaag aagcagtcgc agtgaagatt gtagatatga
     4441 agcgtgccgt agactgtcca gaaaatatta agaaagagat ctgtatcaat aaaatgctaa
     4501 atcatgaaaa tgtagtaaaa ttctatggtc acaggagaga aggcaatatc caatatttat
     4561 ttctggagta ctgtagtgga ggagagcttt ttgacagaat agagccagac ataggcatgc
     4621 ctgaaccaga tgctcagaga ttcttccatc aactcatggc aggggtggtt tatctgcatg
     4681 gtattggaat aactcacagg gatattaaac cagaaaatct tctgttggat gaaagggata
     4741 acctcaaaat ctcagacttt ggcttggcaa cagtatttcg gtataataat cgtgagcgtt
     4801 tgttgaacaa gatgtgtggt actttaccat atgttgctcc agaacttctg aagagaagag
     4861 aatttcatgc agaaccagtt gatgtttggt cctgtggaat agtacttact gcaatgctcg
     4921 ctggagaatt gccatgggac caacccagtg acagctgtca ggagtattct gactggaaag
     4981 aaaaaaaaac atacctcaac ccttggaaaa aaatcgattc tgctcctcta gctctgctgc
     5041 ataaaatctt agttgagaat ccatcagcaa gaattaccat tccagacatc aaaaaagata
     5101 gatggtacaa caaacccctc aagaaagggg caaaaaggcc ccgagtcact tcaggtggtg
     5161 tgtcagagtc tcccagtgga ttttctaagc acattcaatc caatttggac ttctctccag
     5221 taaacagtgc ttctagtgaa gaaaatgtga agtactccag ttctcagcca gaaccccgca
     5281 caggtctttc cttatgggat accagcccct catacattga taaattggta caagggatca
     5341 gcttttccca gcccacatgt cctgatcata tgcttttgaa tagtcagtta cttggcaccc
     5401 caggatcctc acagaacccc tggcagcggt tggtcaaaag aatgacacga ttctttacca
     5461 aattggatgc agacaaatct tatcaatgcc tgaaagagac ttgtgagaag ttgggctatc
     5521 aatggaagaa aagttgtatg aatcaggtta ctatatcaac aactgatagg agaaacaata
     5581 aactcatttt caaagtgaat ttgttagaaa tggatgataa aatattggtt gacttccggc
     5641 tttctaaggg tgatggattg gagttcaaga gacacttcct gaagattaaa gggaagctga
     5701 ttgatattgt gagcagccag aaggtttggc ttcctgccac actcgagggt atggactaca
     5761 aggacgatga cgataagtga gttaacgttt aaacccgctg atcagcctcg actgtgcctt
     5821 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5881 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5941 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6001 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     6061 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6421 cggtctattc ttttgattt
//
