LOCUS       sequence-198188-        8979 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 8979)
  AUTHORS   Kim ST, Xu B, Kastan MB
  TITLE     Involvement of the cohesin protein, Smc1, in Atm-dependent and 
            independent responses to DNA damage.
  JOURNAL   Genes Dev. 2002 Mar 1;16(5):560-70.
  PUBMED    11877376
REFERENCE   2  (bases 1 to 8979)
  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: "Genes 
            Dev."; date: "2002-03-1"; volume: "16(5)"; pages: "560-70"
COMMENT     Sequence Label: pCDNA 3 5' cMyc SMC1 wt
FEATURES             Location/Qualifiers
     source          1..8979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2851..2860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2860..2889
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(6625..6643)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6626..6643)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6663..6680)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6669..6780
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6870..6889)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7080..7101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7220..7240)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7225..7554
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7405..7540
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7467..7486
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7621..8415
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7675..7694)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8285..8304
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8589..8710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8626..8645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8680..8699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8759..8775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8759..8775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8772..8794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8783..8799
                     /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(8807..8813,8814..8831,8832..8837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8852..8873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gacccaagct ccgcgggttt aaacggatcc gccaccatgg aacagaaact catctctgaa
     2881 gaggatctgg aattcatggg gttcctgaaa ctgattgaga ttgagaactt taagtcgtac
     2941 aagggtcgac agattatcgg accatttcag aggttcaccg ccatcattgg acccaatggc
     3001 tctggtaagt caaatctcat ggatgccatc agctttgtgc taggtgaaaa aaccagcaac
     3061 ctgcgggtaa agaccctgcg ggacctgatc catggagctc ctgtgggcaa gccagctgcc
     3121 aaccgggcct ttgtcagcat ggtctactct gaggagggtg ctgaggaccg tacctttgcc
     3181 cgtgtcattg taggaggttc ttctgagtac aagatcaaca acaaagtggt ccaactacat
     3241 gagtacagtg aggaattaga gaagttgggc attctcatca aagctcgtaa cttcctcgtt
     3301 ttccagggtg ctgtggaatc tattgccatg aagaacccca aagagaggac agctctattt
     3361 gaagagatta gtcgttctgg ggagctggcg caggagtatg acaagcgaaa gaaggaaatg
     3421 gtgaaggctg aagaggacac acagtttaat taccatcgca agaaaaatat tgcggctgaa
     3481 cgcaaggaag caaagcagga gaaagaagag gctgaccggt accagcgcct gaaggatgag
     3541 gtagtacggg ctcaggtaca gctgcagctc tttaagcttt accataatga agtggaaatt
     3601 gagaagctca acaaggaact ggcctcaaag aacaaggaga tcgagaagga caagaagcgt
     3661 atggacaagg tggaggatga actgaaggag aagaagaagg agctgggcaa aatgatgcgg
     3721 gagcagcagc agattgagaa ggagatcaag gagaaggact cagaattgaa ccagaagcgg
     3781 cctcagtaca tcaaagccaa ggagaacacc tcccacaaaa tcaagaagct ggaagcagcc
     3841 aagaagtctc tgcagaatgc tcagaagcac tacaagaagc gtaaaggtga catggatgag
     3901 ctggagaagg agatgctgtc agtggagaag gctcggcagg agtttgaaga acggatggaa
     3961 gaagagagtc agagtcaggg cagagatttg acgttggagg agaatcaggt gaagaaatac
     4021 caccggttga aagaagaagc cagcaagaga gcagctaccc tggcccagga gctggagaaa
     4081 ttcaatcgag accagaaagc tgaccaggac cgtctggatc tggaagaacg gaagaaagta
     4141 gagacagagg ccaagatcaa gcaaaagctg cgggaaattg aagagaatca gaagcggatt
     4201 gagaaactgg aggaatacat caccactagc aagcagtccc tagaagagca gaagaagcta
     4261 gagggggagc tgacagagga ggtggagatg gccaagcggc gtattgatga aatcaataag
     4321 gagctgaacc aggtgatgga gcagctaggg gatgcccgca tcgaccgcca ggagagcagc
     4381 cgccagcagc gaaaggcaga gataatggaa agcatcaagc gcctttaccc tggctctgtg
     4441 tacggccgcc tcattgacct atgccagccc acacaaaaga agtatcagat tgctgtaacc
     4501 aaggttttgg gcaagaacat ggatgccatt attgtggact cggagaagac aggccgggac
     4561 tgtattcagt atatcaagga gcagcgtggg gagcctgaga ccttcttgcc tcttgactac
     4621 ctggaggtga agcctacaga tgagaaactc cgggagctga agggggccaa gctagtgatt
     4681 gatgtgattc gctatgagcc acctcatatc aaaaaggccc tgcagtatgc ttgtggcaat
     4741 gcccttgtct gtgacaacgt ggaagatgcc cgccgcattg cctttggagg ccaccagcgc
     4801 cacaagacag tggcactgga tggaacccta ttccagaagt caggagtgat ctctggtggg
     4861 gccagtgacc tgaaggccaa ggcacggcgc tgggatgaga aagcagtaga caagttgaaa
     4921 gagaagaagg agcgcttgac agaggagctg aaagagcaga tgaaggcaaa acggaaagag
     4981 gcagagctgc gtcaggtgca gtctcaggcc catggactgc agatgcggct caagtactcc
     5041 cagagtgacc tagaacagac caagacacga catctagccc tgaatctgca ggaaaaatcc
     5101 aagctggaga gtgagctagc caactttggg cctcgcatta atgatatcaa gaggatcatt
     5161 cagagccgag agagggaaat gaaagacttg aaggagaaga tgaaccaggt agaggatgag
     5221 gtgtttgaag agttttgtcg ggagattggt gtgcgcaaca tccgggagtt tgaggaagaa
     5281 aaggtgaaac ggcagaatga aatcgccaag aagcgtttgg agtttgagaa tcagaagact
     5341 cgcttgggca ttcagttgga ttttgaaaag aaccaactga aggaggacca agataaagta
     5401 cacatgtggg agcagacagt gaaaaaagat gaaaatgaga tagaaaagct caaaaaggag
     5461 gaacaaagac acatgaagat catagatgag accatggctc agctacaaga cctgaagaat
     5521 cagcatctgg ccaagaagtc ggaagtgaat gacaagaatc atgagatgga ggagattcgt
     5581 aagaaactcg ggggcgccaa caaggaaatg acccatttac agaaggaggt gacagccatt
     5641 gagaccaagc ttgaacagaa gcgcagtgac cgtcacaact tgctacaggc ctgtaagatg
     5701 caggacatta agttgccact gtcaaaaggc accatggatg atattagtca ggaagagggt
     5761 agctcccagg gggaggactc agtgagtggt tcacagagaa tttccagtat ctatgcacga
     5821 gaggccctca ttgagattga ctacggtgat ctgtgtgagg atctgaagga tgcccaggct
     5881 gaggaagaga tcaagcaaga gatgaacaca ctgcagcaga agctgaatga gcagcagagt
     5941 gtgcttcagc gtattgccgc ccccaacatg aaggccatgg aaaagctgga aagtgtccga
     6001 gacaagttcc aggagacctc agatgagttt gaagcagccc gaaagcgagc aaagaaggcc
     6061 aagcaggcat tcgaacagat caagaaggag cgctttgacc gcttcaatgc ttgttttgaa
     6121 tctgtggcta ccaacattga tgagatctat aaggccctgt cccgcaatag cagtgcccag
     6181 gcattcctgg gccctgagaa ccctgaagag ccctacttgg atggcatcaa ctacaactgt
     6241 gtggctcctg ggaaacgctt ccggcctatg gacaacttgt caggcgggga gaagacagtg
     6301 gcagctctgg ccctgctctt tgccatccac agctacaagc cagccccctt cttcgtcctg
     6361 gatgagattg atgctgcctt ggataacacc aacattggca aggtggcaaa ttacatcaag
     6421 gagcagtcga cttgcaactt ccaggccatc gtcatctctc tcaaggagga gttctacacc
     6481 aaggccgaga gcctcattgg agtctatcct gagcaagggg actgtgtgat cagcaaagtc
     6541 ctgaccttcg acctcaccaa gtacccagat gccaacccca accccaatga gcagtagctc
     6601 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     6661 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6721 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6781 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcmgs rcagcgtgrc
     6841 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6901 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6961 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     7021 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     7081 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     7141 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     7201 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     7261 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     7321 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7381 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     7441 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     7501 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     7561 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     7621 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     7681 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     7741 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     7801 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     7861 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     7921 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     7981 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     8041 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     8101 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     8161 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     8221 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     8281 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     8341 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     8401 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     8461 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     8521 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     8581 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     8641 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     8701 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     8761 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     8821 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     8881 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     8941 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct
//
