LOCUS       sequence-451632-        9306 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 9306)
  AUTHORS   Duxin JP, Moore HR, Sidorova J, Karanja K, Honaker Y, Dao B, 
            Piwnica-Worms H, Campbell JL, Monnat RJ Jr, Stewart SA
  TITLE     Okazaki Fragment Processing-independent Role for Human Dna2 Enzyme 
            during DNA Replication.
  JOURNAL   J Biol Chem. 2012 Jun 22;287(26):21980-91. Epub 2012 May 7.
  PUBMED    22570476
REFERENCE   2  (bases 1 to 9306)
  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: "J Biol 
            Chem."; date: "2012-06-22"; volume: "287(26)"; pages: "21980-91. 
            Epub 2012 May 7"
COMMENT     Sequence Label: pBabe hygro 3xFLAG Dna2 wt
FEATURES             Location/Qualifiers
     source          1..9306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      431..440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3703..3723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3708..4037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3888..4023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4111..5136
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4111..5136
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             5934..6403
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6533..6550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6704..7292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7452..8243,8244..8312))
                     /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     8075..8094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8313..8417)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8907..9106
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc
      421 cggatccgcc gccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat
      481 cgattacaag gatgacgatg acttaattaa agagcagctg aacgaactgg agctgctgat
      541 ggagaagagt ttttgggagg aggcggagct gccggcggag ctatttcaga agaaagtggt
      601 agcttccttt ccaagaacag ttctgagcac aggaatggat aaccggtacc tggtgttggc
      661 agtcaatact gtacagaaca aagagggaaa ctgtgaaaag cgcctggtca tcactgcttc
      721 acagtcacta gaaaataaag aactatgcat ccttaggaat gactggtgtt ctgttccagt
      781 agagccagga gatatcattc atttggaggg agactgcaca tctgacactt ggataataga
      841 taaagatttt ggatatttga ttctgtatcc agacatgctg atttctggca ccagcatagc
      901 cagtagtatt cgatgtatga gaagagctgt cctgagtgaa acttttagga gctctgatcc
      961 agccacacgc caaatgctaa ttggtacggt tctccatgag gtgtttcaaa aagccataaa
     1021 taatagcttt gccccagaaa agctacaaga acttgctttt caaacaattc aagaaataag
     1081 acatttgaag gaaatgtacc gcttaaatct aagtcaagat gaaataaaac aagaagtaga
     1141 ggactatctt ccttcgtttt gtaaatgggc aggagatttc atgcataaaa acacttcgac
     1201 tgacttccct cagatgcagc tctctctgcc aagtgataat agtaaggata attcaacatg
     1261 taacattgaa gtcgtgaaac caatggatat tgaagaaagc atttggtccc ctaggtttgg
     1321 attgaaaggc aaaatagatg ttacagttgg tgtgaaaata catcgagggt ataaaacaaa
     1381 atacaagata atgccgctgg aacttaaaac tggcaaagaa tcaaattcta ttgaacaccg
     1441 tagtcaggtt gttctgtaca ctctactaag ccaagagaga agagctgatc cagaggctgg
     1501 cttgcttctc tacctcaaga ctggtcagat gtaccctgtg cctgccaacc atctagataa
     1561 aagagaatta ttaaagctaa gaaaccagat ggcattctca ttgtttcacc gtattagcaa
     1621 atctgctact agacagaaga cacagcttgc ttctttgcca caaataattg aggaagagaa
     1681 aacttgtaaa tattgttcac aaattggcaa ttgtgctctt tatagcagag cagttgaaca
     1741 acagatggat tgtagttcag tcccaattgt gatgctgccc aaaatagaag aagaaaccca
     1801 gcatctgaag caaacacact tagaatattt cagcctttgg tgtctaatgt taaccctgga
     1861 gtcacaatcg aaggataata aaaagaatca ccaaaatatc tggctaatgc ctgcttcgga
     1921 aatggagaag agtggcagtt gcattggaaa cctgattaga atggaacatg taaagatagt
     1981 ttgtgatggg caatatttac ataatttcca atgtaaacat ggtgccatac ctgtcacaaa
     2041 tctaatggca ggtgacagag ttattgtaag tggagaagaa aggtcactgt ttgctttgtc
     2101 tagaggatat gtgaaggaga ttaacatgac aacagtaact tgtttattag acagaaactt
     2161 gtcggtcctt ccagaatcaa ctttgttcag attagaccaa gaagaaaaaa attgtgatat
     2221 agatacccca ttaggaaatc tttccaaatt gatggaaaac acgtttgtca gcaaaaaact
     2281 tcgagattta attattgact ttcgtgaacc tcagtttata tcctacctta gttctgttct
     2341 tccacatgat gcaaaggata cagttgcctg cattctaaag ggtttgaata agcctcagag
     2401 gcaagcgatg aaaaaggtac ttctttcaaa agactacaca ctcatcgtgg gtatgcctgg
     2461 gacaggaaaa acaactacga tatgtactct cgtaagaatt ctctacgcct gtggttttag
     2521 cgttttgttg accagctata cacactctgc tgttgacaat attcttttga agttagccaa
     2581 gtttaaaata ggatttttgc gtttgggtca gattcagaag gttcatccag ctatccagca
     2641 atttacagag caagaaattt gcagatcaaa gtccattaaa tccttagctc ttctagaaga
     2701 actctacaat agtcaactta tagttgcaac aacatgtatg ggaataaacc atccaatatt
     2761 ttcccgtaaa atttttgatt tttgtattgt ggatgaagcc tctcaaatta gccaaccaat
     2821 ttgtctgggc cccctttttt tttcacggag atttgtgtta gtgggggacc atcagcagct
     2881 tcctcccctg gtgctaaacc gtgaagcaag agctcttggc atgagtgaaa gcttattcaa
     2941 gaggctggag cagaataaga gtgctgttgt acagttaacc gtgcagtaca gaatgaacag
     3001 taaaattatg tccttaagta ataagctgac ctatgagggc aagctggagt gtggatcaga
     3061 caaagtggcc aatgcagtga taaacctacg tcactttaaa gatgtgaagc tggaactgga
     3121 attttatgct gactattctg ataatccttg gttgatggga gtatttgaac ccaacaatcc
     3181 tgtttgtttc cttaatacag acaaggttcc agcgccagaa caagttgaaa aaggtggtgt
     3241 gagcaatgta acagaagcca aactcatagt tttcctaacc tccatttttg ttaaggctgg
     3301 atgcagtccc tctgatattg gtattattgc accgtacagg cagcaattaa agatcatcaa
     3361 tgatttattg gcacgttcta ttgggatggt cgaagttaat acagtagaca aataccaagg
     3421 aagggacaaa agtattgtcc tagtatcttt tgttagaagt aataaggatg gaactgttgg
     3481 tgaactcttg aaagattggc gacgtcttaa tgttgctata accagagcca aacataaact
     3541 gattcttctg gggtgtgtgc cctcactaaa ttgctatcct cctttggaga agctgcttaa
     3601 tcatttaaac tcagaaaaat taatcattga tcttccatca agagaacatg aaagtctttg
     3661 ccacatattg ggtgactttc aaagagaata ggtcgaccct gtggaatgtg tgtcagttag
     3721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3961 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4021 gcctaggctt ttgcaaaaag cttgccagct ggggcgccct ctggtaaggt tgggaagccc
     4081 tgcagatctg atcccggggg gcaatgagat atgaaaaagc ctgaactcac cgcgacgtct
     4141 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     4201 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     4261 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     4321 gcgctcccga ttccggaagt gcttgacatt ggggaattta gcgagagcct gacctattgc
     4381 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     4441 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     4501 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     4561 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     4621 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     4681 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     4741 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     4801 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     4861 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     4921 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     4981 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     5041 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     5101 cgacgcccca gcactcgtcc gagggcaaag gaatagagta gatgccgacc gggatctatc
     5161 gatgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5221 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5641 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5701 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgtaa ccaccacacc
     5761 cgccgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     5821 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     5881 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     5941 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6001 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6061 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     6121 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     6181 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     6241 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     6301 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     6361 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     6421 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     6481 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6541 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6601 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6661 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6721 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6781 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6841 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6901 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6961 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7021 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7081 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7141 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7201 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7261 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7321 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7381 ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact
     7441 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7501 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7561 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7621 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7681 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7741 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7801 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7861 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7921 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7981 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8041 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8101 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8161 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8221 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8281 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8341 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8401 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc
     8461 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     8521 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     8581 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     8641 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     8701 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     8761 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     8821 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
     8881 ggaccaccga cccaccaccg ggaggcaagc tggccagcaa cttatctgtg tctgtccgat
     8941 tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc
     9001 tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg
     9061 agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg
     9121 tggaatccga ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc
     9181 ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg
     9241 cagcgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa
     9301 aattag
//
