LOCUS       sequence-190209-        5832 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 5832)
  AUTHORS   Tang ED, Nunez G, Barr FG, Guan KL
  TITLE     Negative regulation of the forkhead transcription factor FKHR by 
            Akt.
  JOURNAL   J Biol Chem. 1999 Jun 11. 274(24):16741-6.
  PUBMED    10358014
REFERENCE   2  (bases 1 to 5832)
  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. 1999 Jun 11. 274(24):16741-6."
COMMENT     Sequence Label: 3xIRS luciferase
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    422..556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(698..715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(869..1457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(949..968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1628..2419,2420..2488))
                     /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     2251..2270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2489..2593)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2620..3075
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2707..2726)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2917..2938
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3265..3386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3495..3691
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3542..3677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3721..5373
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(3787..3805)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          5614..5679
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5809..5829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat
       61 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag
      121 aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac
      181 atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa
      241 aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat
      301 agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt
      361 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
      421 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
      481 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
      541 ttatcatgtc tggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
      601 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
      661 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
      721 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      781 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      841 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      901 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      961 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1021 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1081 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1141 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1201 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1261 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1321 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1381 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1441 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1501 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1561 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1621 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1681 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1741 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1801 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1861 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1921 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1981 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2041 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2101 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2161 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2221 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2281 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2341 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2401 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2461 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2521 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2581 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     2641 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2701 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2761 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2821 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2881 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2941 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3001 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3061 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3121 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     3181 tacgtggagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3241 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3301 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3361 ttgtccaaac tcatcaatgt atcttatggt actgtaactg agctaacata acccgggagg
     3421 tacgcaaaac aaacttattt tgaagcaaaa caaacttatt ttgaagcaaa acaaacttat
     3481 tttgaatcga gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3541 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3601 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3661 aggctttttt ggaggcctag gcttttgcaa aaagcttggc attccggtac tgttggtaaa
     3721 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatcctct agaggatgga
     3781 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
     3841 gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc
     3901 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
     3961 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
     4021 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgaacatt
     4081 tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     4141 aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga
     4201 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     4261 tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa ttcctctgga
     4321 tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg
     4381 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     4441 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     4501 cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct tcaggattac
     4561 aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg
     4621 attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg
     4681 aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat
     4741 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     4801 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     4861 acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt
     4921 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     4981 ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg cttgaagtct
     5041 ttaattaaat acaaaggata tcaggtggcc cccgctgaat tggaatcgat attgttacaa
     5101 caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc cggtgaactt
     5161 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     5221 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     5281 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     5341 aaggccaaga agggcggaaa gtccaaattg taaaatgtaa ctgtattcag cgatgacgaa
     5401 attcttagct attgtaatac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca
     5461 gttattggtg cccttaaacg cctggttgct acgcctgaat aagtgataat aagcggatga
     5521 atggcagaaa ttcgccggat ctttgtgaag gaaccttact tctgtggtgt gacataattg
     5581 gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa
     5641 tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat
     5701 gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc agaagaaatg
     5761 ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag
     5821 agaaaggtag aa
//
