LOCUS       sequence-6922-ex        5355 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 5355)
  AUTHORS   Newman RM, Mobascher A, Mangold U, Koike C, Diah S, Schmidt M, 
            Finley D, Zetter BR
  TITLE     Antizyme targets cyclin D1 for degradation. A novel mechanism for 
            cell growth repression.
  JOURNAL   J Biol Chem. 2004 Oct 1. 279(40):41504-11.
  PUBMED    15277517
REFERENCE   2  (bases 1 to 5355)
  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. 2004 Oct 1. 279(40):41504-11."
COMMENT     Sequence Label: pFLAG rat Antizyme 1
FEATURES             Location/Qualifiers
     source          1..5355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1696..2318
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1895..1914)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2342..2362)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2347..2676
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2527..2662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2714..2733)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2715..2733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2747..2763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2747..2763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2760..2782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2771..2787
                     /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(2795..2801,2802..2819,2820..2825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2840..2861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2978..2995)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3149..3737)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3229..3248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3908..4699,4700..4768))
                     /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     4531..4550
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4769..4873)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4900..5328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4987..5006)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5197..5218
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tatggtgaaa tcctccctgc agcggatcct caacagccac tgcttcgcca gagagaaaga
     1021 aggggacaaa cgcagcgcca cgcttcacgc cagccgcacc atgccgcttc ttagtcagca
     1081 cagccgcggc ggctgcagca gcgagagttc tagggttgcc cttcattgct gtagtaacct
     1141 gggtccgggg cctcggtggt gctccgatgt ccctcaccca cccctgaaga tcccaggtgg
     1201 gcgagggaac agtcagcggg atcacagtct ttcagctagc atcctgtact ccgacgagcg
     1261 gctgaatgtg acagaggagc cgacgtctaa cgacaagacc agggtcctga gcatccagtg
     1321 cacgctcacg gaggccaagc aggtcacctg gagggcggtg tggaatggtg gtggcctcta
     1381 catcgagctc ccagctgggc ctctgccaga aggcagcaag gacagttttg cagctctact
     1441 agagttcgca gaggagcagc tccgggccga ccacgttttc atctgcttcc ccaagaaccg
     1501 tgaggacaga gccgccctac tccgaacctt cagctttctt ggctttgaga ttgtgagacc
     1561 cgggcatccc ctcgtcccca agagacccga cgcttgcttc atggtctaca cgctggagag
     1621 agaggacccg ggtgaggagg actaggaatt catcgataga tctgatatcg gtaccagtcg
     1681 actctagagg atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     1741 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     1801 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     1861 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     1921 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     1981 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     2041 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     2101 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     2161 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     2221 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     2281 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg
     2341 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2401 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2461 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2521 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2581 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2641 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcctcgagga actgaaaaac
     2701 cagaaagtta attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc
     2761 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2821 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2881 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2941 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3001 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3061 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3301 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3361 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3421 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3481 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3541 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3601 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3661 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3721 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3901 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3961 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4021 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4081 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4141 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4201 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4261 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4321 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4381 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4441 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4501 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4561 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4621 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4681 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4741 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4801 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4861 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4921 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4981 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5041 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5101 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5161 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5221 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5281 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5341 taacgcttac aattt
//
