LOCUS       sequence-258313-        6766 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 6766)
  AUTHORS   Hinds PW, Mittnacht S, Dulic V, Arnold A, Reed SI, Weinberg RA
  TITLE     Regulation of retinoblastoma protein functions by ectopic expression
            of human cyclins.
  JOURNAL   Cell. 1992 Sep 18. 70(6):993-1006.
  PUBMED    1388095
REFERENCE   2  (bases 1 to 6766)
  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: "Cell. 1992 
            Sep 18. 70(6):993-1006."
COMMENT     Sequence Label: Rc/CMV cyclin D3
FEATURES             Location/Qualifiers
     source          1..6766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2227..2245)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2228..2245)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2265..2282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2271..2495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2541..3054
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2628..2647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2838..2859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3077..3097)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3082..3411
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3262..3397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3478..4272
                     /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(3532..3551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4142..4161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4446..4580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4483..4502)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4537..4556
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4611..4627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4611..4627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4624..4646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4635..4651
                     /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(4659..4665,4666..4683,4684..4689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4704..4725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4842..4859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5013..5601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5093..5112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5772..6563,6564..6632))
                     /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     6395..6414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6633..6737)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
      901 ccgagctcgg atccactagt aacggccgcc aaattcgggg ccagcgccag cgccagaccc
      961 gcgccccgcg ctctccggcc cgtcgcctgc cttgggactc gcgagcccgc actcccgccc
     1021 tgcctgttcg ctgcccgagt atggagctgc tgtgttgcga aggcacccgg cacgcgcccc
     1081 gggccgggcc ggacccgcgg ctgctggggg accagcgtgt cctgcagagc ctgctccgcc
     1141 tggaggagcg ctacgtaccc cgcgcctcct acttccagtg cgtgcagcgg gagatcaagc
     1201 cgcacatgcg gaagatgctg gcttactgga tgctggaggt atgtgaggag cagcgctgtg
     1261 aggaggaagt cttccccctg gccatgaact acctggatcg ctacctgtct tgcgtcccca
     1321 cccgaaaggc gcagttgcag ctcctgggtg cggtctgcat gctgctggcc tccaagctgc
     1381 gcgagaccac gcccctgacc atcgaaaaac tgtgcatcta caccgaccac gctgtctctc
     1441 cccgccagtt gcgggactgg gaggtgctgg tcctagggaa gctcaagtgg gacctggctg
     1501 ctgtgattgc acatgatttc ctggccttca ttctgcaccg gctctctctg ccccgtgacc
     1561 gacaggcctt ggtcaaaaag catgcccaga cctttttggc cctctgtgct acagattata
     1621 cctttgccat gtacccgcca tccatgatcg ccacgggcag cattggggct gcagtgcaag
     1681 gcctgggtgc ctgctccatg tccggggatg agctcacaga gctgctggca gggatcactg
     1741 gcactgaagt ggactgcctg cgggcctgtc aggagcagat cgaagctgca ctcagggaga
     1801 gcctcaggga agcctctcag accagctcca gcccagcgcc caaagccccc cggggctcca
     1861 gcagccaagg gcccagccag accagcactc ctacagatgt cacagccata cacctgtagc
     1921 cctggagagg ccctctggag tggccaagga gctgctatag cctggggtgg ggtcatgccc
     1981 tcctctccca ttgtccctct gccccatcct ccagcaggga aaatgcagca gggatgccct
     2041 ggaggtggct gagcccctgt ctagagaggg aggcaagccc tgttgacaca ggtctttcct
     2101 aaggctgcaa ggtttaggct ggtggcccag gaccatcatc ctactgtaat aaagatgatt
     2161 gtgaaataaa actggctttg gcccgaattc tggcggccgc tcgagcatgc atctagaggg
     2221 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     2281 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2341 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2401 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2461 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2521 gggctcgagg ggggatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2881 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2941 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaaat
     3001 atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg gggtgggtac
     3061 cgagctcgaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3241 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3301 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3361 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3421 ggagcttgga tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3481 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3541 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3601 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3661 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3721 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3781 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3841 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3901 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3961 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4021 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4081 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4141 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4201 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4261 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4321 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4381 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4441 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4561 tatcttatca tgtctggatc cgtcgacctc gagagcttgg cgtaatcatg gtcatagctg
     4621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacg
//
