LOCUS       sequence-178398-        6778 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 6778)
  AUTHORS   Watanabe H, Pan ZQ, Schreiber-Agus N, DePinho RA, Hurwitz J, Xiong Y
  TITLE     Suppression of cell transformation by the cyclin-dependent kinase 
            inhibitor p57KIP2 requires binding to proliferating cell nuclear 
            antigen.
  JOURNAL   Proc Natl Acad Sci U S A. 1998 Feb 17. 95(4):1392-7.
  PUBMED    9465025
REFERENCE   2  (bases 1 to 6778)
  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: "Proc Natl 
            Acad Sci U S A. 1998 Feb 17. 95(4):1392-7."
COMMENT     Sequence Label: pCMV-Cyclin D1
FEATURES             Location/Qualifiers
     source          1..6778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(130..148)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(131..148)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(168..185)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    174..398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      444..872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(531..550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     741..762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(881..901)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        886..1215
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1066..1201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1128..1147
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1282..2076
                     /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(1336..1355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1946..1965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2250..2371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2287..2306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2341..2360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2420..2436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2420..2436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2433..2455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2444..2460
                     /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(2468..2474,2475..2492,2493..2498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2513..2534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2651..2668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2822..3410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2902..2921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3581..4372,4373..4441))
                     /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     4204..4223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4442..4546)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4621..4640)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4812..5191
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5192..5395
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5346..5366
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5440..5459
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5440..5458
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6268..6277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
       61 aaaaaaaagg aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga
      121 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
      181 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
      241 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
      301 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
      361 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
      421 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
      481 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      541 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      601 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      661 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      721 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
      781 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
      841 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
      901 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      961 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1021 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     1081 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1141 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     1201 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     1261 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     1321 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     1381 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     1441 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     1501 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     1561 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     1621 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     1681 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     1741 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     1801 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     1861 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     1921 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     1981 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     2041 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     2101 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     2161 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     2221 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     2281 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2341 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     2401 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2461 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2521 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2581 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4561 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     4621 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     4681 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     4741 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     4801 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4861 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4921 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4981 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5041 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5101 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5161 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     5221 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     5281 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5341 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     5401 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     5461 gacccaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc
     5521 ccgcagtagc agcgagcagc agagtccgca cgctccggcg aggggcagaa gagcgcgagg
     5581 gagcgcgggg cagcagaagc gagagccgag cgcggaccca gccaggaccc acagccctcc
     5641 ccagctgccc aggaagagcc ccagccatgg aacaccagct cctgtgctgc gaagtggaaa
     5701 ccatccgccg cgcgtacccc gatgccaacc tcctcaacga ccgggtgctg cgggccatgc
     5761 tgaaggcgga ggagacctgc gcgccctcgg tgtcctactt caaatgtgtg cagaaggagg
     5821 tcctgccgtc catgcggaag atcgtcgcca cctggatgct ggaggtctgc gaggaacaga
     5881 agtgcgagga ggaggtcttc ccgctggcca tgaactacct ggaccgcttc ctgtcgctgg
     5941 agcccgtgaa aaagagccgc ctgcagctgc tgggggccac ttgcatgttc gtggcctcta
     6001 agatgaagga gaccatcccc ctgacggccg agaagctgtg catctacacc gacaactcca
     6061 tccggcccga ggagctgctg caaatggagc tgctcctggt gaacaagctc aagtggaacc
     6121 tggccgcaat gaccccgcac gatttcattg aacacttcct ctccaaaatg ccagaggcgg
     6181 aggagaacaa acagatcatc cgcaaacacg cgcagacctt cgttgccctc tgtgccacag
     6241 atgtgaagtt catttccaat ccgccctcca tggtggcagc ggggagcgtg gtggccgcag
     6301 tgcaaggcct gaacctgagg agccccaaca acttcctgtc ctactaccgc ctcacacgct
     6361 tcctctccag agtgatcaag tgtgacccag actgcctccg ggcctgccag gagcagatcg
     6421 aagccctgct ggagtcaagc ctgcgccagg cccagcagaa catggacccc aaggccgccg
     6481 aggaggagga agaggaggag gaggaggtgg acctggcttg cacacccacc gacgtgcggg
     6541 acgtggacat ctgagggcgc caggcaggcg ggcgccaccg ccacccgcag cgagggcgga
     6601 gccggcccca ggtgctccac tgacagtccc tcctctccgg agcattttga taccagaagg
     6661 gaaagcttca ttctccttgt tgttggttgt tttttccttt gctctttccc ccttccatct
     6721 ctgacttaag caaaagaaaa agattaccca aaaactgtct ttaaaagaga gagagaga
//
