LOCUS       sequence-231060-        6619 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 6619)
  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 6619)
  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: pcDNA cyclin D1 HA T286A
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1073..1082
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1360..1386
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1464..1482)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1465..1482)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1502..1519)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1508..1732
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1778..2291
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1865..1884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2075..2096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2314..2334)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2319..2648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2499..2634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2715..3509
                     /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(2769..2788)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3379..3398
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3683..3817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3720..3739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3774..3793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3848..3864)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3848..3864)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3861..3883)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3872..3888
                     /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(3896..3902,3903..3920,3921..3926))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3941..3962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4079..4096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4250..4838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4330..4349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5009..5800,5801..5869))
                     /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     5632..5651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5870..5974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6049..6068)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6240..6619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac
      301 tagtaacggc cgccagaatt cgggggcgca gtagcagcga gcagcagagt ccgcacgctc
      361 cggcgagggg cagaagagcg cgagggagcg cggggcagca gaagcgagag ccgagcgcgg
      421 acccagccag gacccacagc cctccccagc tgcccaggaa gagccccagc catggaacac
      481 cagctcctgt gctgcgaagt ggaaaccatc cgccgcgcgt accccgatgc caacctcctc
      541 aacgaccggg tgctgcgggc catgctgaag gcggaggaga cctgcgcgcc ctcggtgtcc
      601 tacttcaaat gtgtgcagaa ggaggtcctg ccgtccatgc ggaagatcgt cgccacctgg
      661 atgctggagg tctgcgagga acagaagtgc gaggaggagg tcttcccgct ggccatgaac
      721 tacctggacc gcttcctgtc gctggagccc gtgaaaaaga gccgcctgca gctgctgggg
      781 gccacttgca tgttcgtggc ctctaagatg aaggagacca tccccctgac ggccgagaag
      841 ctgtgcatct acaccgacaa ctccatccgg cccgaggagc tgctgcaaat ggagctgctc
      901 ctggtgaaca agctcaagtg gaacctggcc gcaatgaccc cgcacgattt cattgaacac
      961 ttcctctcca aaatgccaga ggcggaggag aacaaacaga tcatccgcaa acacgcgcag
     1021 accttcgttg ccctctgtgc cacagatgtg aagttcattt ccaatccgcc ctccatggtg
     1081 gcagcgggga gcgtggtggc cgcagtgcaa ggcctgaacc tgaggagccc caacaacttc
     1141 ctgtcctact accgcctcac acgcttcctc tccagagtga tcaagtgtga cccagactgc
     1201 ctccgggcct gccaggagca gatcgaagct ctgctggagt caagcctgcg ccaggcccag
     1261 cagaacatgg accccaaggc cgccgaggag gaggaagagg aggaggagga ggtggacctg
     1321 gcttgcgcac ccaccgacgt gcgggacgtg gacatcggct acccctacga cgtcccggac
     1381 tacgccgata tctgatccgg agcattttga taccagaagg gaaagcttgg cggccgctcg
     1441 agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     1501 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1561 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1621 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1681 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1741 gcggaaagaa ccagctgggg ctcgaggggg gatccccacg cgccctgtag cggcgcatta
     1801 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1861 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1921 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1981 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2041 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2101 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2161 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2221 acgcttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct tttctgatta
     2281 tcaaccgggg tgggtaccga gctcgaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2401 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2461 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2521 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2581 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2641 tttgcaaaaa gctcccggga gcttggatat ccattttcgg atctgatcaa gagacaggat
     2701 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2761 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2821 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2881 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2941 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3001 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3061 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3121 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3181 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3241 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3301 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3361 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3421 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3481 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3541 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3601 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3661 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     3721 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3781 gtccaaactc atcaatgtat cttatcatgt ctggatccgt cgacctcgag agcttggcgt
     3841 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3901 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3961 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4981 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5041 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5101 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5161 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5221 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5281 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5341 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5401 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5461 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5521 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5581 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5641 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5701 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5761 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5821 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5881 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5941 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6001 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     6061 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     6241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     6301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6601 agtcatcgct attaccatg
//
