LOCUS       sequence-175310-        7378 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 7378)
  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 7378)
  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 A
FEATURES             Location/Qualifiers
     source          1..7378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(52..70)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(53..70)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(90..107)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    96..320
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      366..879
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(453..472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     663..684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(902..922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        907..1236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1087..1222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1303..2097
                     /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(1357..1376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1967..1986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2271..2405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2308..2327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2362..2381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2436..2452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2436..2452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2449..2471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2460..2476
                     /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(2484..2490,2491..2508,2509..2514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2529..2550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2667..2684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2838..3426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2918..2937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3597..4388,4389..4457))
                     /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     4220..4239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4458..4562)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4637..4656)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4828..5207
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5208..5411
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5362..5382
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5456..5475
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5456..5474
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cccgaattca gatcctggcg gccgctcgag catgcatcta gagggcccta ttctatagtg
       61 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
      121 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
      181 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
      241 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
      301 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgagggggga
      361 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      421 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      481 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      541 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
      601 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      661 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      721 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      781 atttaacgcg aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat
      841 cttcctgttt ttggggcttt tctgattatc aaccggggtg ggtaccgagc tcgaattctg
      901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     1021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     1201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttggatatcc
     1261 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     1321 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     1381 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     1441 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     1501 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     1561 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     1621 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     1681 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     1741 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     1801 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     1861 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     1921 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     1981 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     2041 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     2101 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     2161 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     2221 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     2281 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2341 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2401 ggatccgtcg acctcgagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2461 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2521 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2581 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2641 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2701 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2761 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2821 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2881 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2941 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3001 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3061 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3121 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3181 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3241 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3301 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3361 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3481 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3541 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3601 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3661 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3721 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3781 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3841 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3901 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3961 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4021 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4081 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4141 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4201 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4261 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4321 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4381 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4441 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4501 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4561 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     4621 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     4681 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     4741 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     4801 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     4861 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4921 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4981 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     5041 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     5101 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     5161 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     5221 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     5281 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     5341 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     5401 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
     5461 cgactcacta tagggagacc caagcttggt accgagctcg gatccactag taacggccgc
     5521 cagatctgaa ttcgctgcat ctctgggcgt ctttggctcg ccacgctggg cagtgcctgc
     5581 ctgcgccttt cgcaacctcc tcggccctgc gtggtctcga gctgggtgag cgagcgggcg
     5641 ggctggtagg ctggcctggg ctgcgaccgg cggctacgac tattctttgg ccgggtcggt
     5701 gcgagtggtc ggctgggcag agtgcacgct gcttggcgcc gcaggctgat cccgccgtcc
     5761 actcccggga gcagtgatgt tgggcaactc tgcgccgggg cctgcgaccc gcgaggcggg
     5821 ctcggcgctg ctagcattgc agcagacggc gctccaagag gaccaggaga atatcaaccc
     5881 ggaaaaggca gcgcccgtcc aacaaccgcg gacccgggcc gcgctggcgg tactgaagtc
     5941 cgggaacccg cggggtctag cgcagcagca gaggccgaag acgagacggg ttgcacccct
     6001 taaggatctt cctgtaaatg atgagcatgt caccgttcct ccttggaaag caaacagtaa
     6061 acagcctgcg ttcaccattc atgtggatga agcagaaaaa gaagctcaga agaagccagc
     6121 tgaatctcaa aaaatagagc gtgaagatgc cctggctttt aattcagcca ttagtttacc
     6181 tggacccaga aaaccattgg tccctcttga ttatccaatg gatggtagtt ttgagtcacc
     6241 acatactatg gacatgtcaa ttgtattaga agatgaaaag ccagtgagtg ttaatgaagt
     6301 accagactac catgaggata ttcacacata ccttagggaa atggaggtta aatgtaaacc
     6361 taaagtgggt tacatgaaga aacagccaga catcactaac agtatgagag ctatcctcgt
     6421 ggactggtta gttgaagtag gagaagaata taaactacag aatgagaccc tgcatttggc
     6481 tgtgaactac attgataggt tcctgtcttc catgtcagtg ctgagaggaa aacttcagct
     6541 tgtgggcact gctgctatgc tgttagcctc aaagtttgaa gaaatatacc ccccagaagt
     6601 agcagagttt gtgtacatta cagatgatac ctacaccaag aaacaagttc tgagaatgga
     6661 gcatctagtt ttgaaagtcc ttacttttga cttagctgct ccaacagtaa atcagtttct
     6721 tacccaatac tttctgcatc agcagcctgc aaactgcaaa gttgaaagtt tagcaatgtt
     6781 tttgggagaa ttaagtttga tagatgctga cccatacctc aagtatttgc catcagttat
     6841 tgctggagct gcctttcatt tagcactcta cacagtcacg ggacaaagct ggcctgaatc
     6901 attaatacga aagactggat ataccctgga aagtcttaag ccttgtctca tggaccttca
     6961 ccagacctac ctcaaagcac cacagcatgc acaacagtca ataagagaaa agtacaaaaa
     7021 ttcaaagtat catggtgttt ctctcctcaa cccaccagag acactaaatc tgtaacaatg
     7081 aaagactgcc tttgttttct aagatgtaaa tcactcaaag tatatggtgt acagttttta
     7141 acttaggttt taattttaca atcatttctg aatacagaag ttgtggccaa gtacaaatta
     7201 tggtatctat tactttttaa atggttttaa tttgtatatc ttttgtatat gtatctgtct
     7261 tagatatttg gctaatttta agtggttttg ttaaagtatt aatgatgcca gctgtcagga
     7321 taataaattg atttggaaaa aaaaaaaaaa aaaaanccgt cgacaaagcg gccgcaaa
//
