LOCUS       sequence-132751-        6618 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 6618)
  AUTHORS   Baker GL, Landis MW, Hinds PW
  TITLE     Multiple functions of D-type cyclins can antagonize pRb-mediated 
            suppression of proliferation.
  JOURNAL   Cell Cycle. 2005 Feb . 4(2):330-8.
  PUBMED    15684604
REFERENCE   2  (bases 1 to 6618)
  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 Cycle.
            2005 Feb . 4(2):330-8."
COMMENT     Sequence Label: Rc/CMV cyclin D1 K112E HA
FEATURES             Location/Qualifiers
     source          1..6618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    252..386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(276..295)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     330..349
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(560..1354)
                     /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(671..690)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1281..1300
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1421..1750)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1435..1570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1735..1755
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1778..2291)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1973..1994)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2185..2204
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2337..2561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2550..2567
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2587..2605
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2587..2604
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2683..2709)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2987..2996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3800..3819)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3801..3819)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3864..4067)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3893..3913)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4068..4447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4619..4638
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4713..4817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4818..4886,4887..5678)
                     /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     complement(5036..5055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5849..6437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6338..6357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6591..6608
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 cgaggtcgac ggatccagac atgataagat acattgatga gtttggacaa accacaacta
      301 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      361 ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg
      421 cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag
      481 aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt
      541 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
      601 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
      661 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
      721 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
      781 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
      841 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
      901 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
      961 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     1021 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     1081 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     1141 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     1201 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     1261 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     1321 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     1381 gatcagatcc gaaaatggat atccaagctc ccgggagctt tttgcaaaag cctaggcctc
     1441 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     1501 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     1561 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     1621 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     1681 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     1741 actgacacac attccacaga attcgagctc ggtacccacc ccggttgata atcagaaaag
     1801 ccccaaaaac aggaagattg tataagcaaa tatttaaatt gtaagcgtta atattttgtt
     1861 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     1921 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     1981 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2041 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2101 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2161 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2221 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2281 acagggcgcg tggggatccc ccctcgagcc ccagctggtt ctttccgcct cagaagccat
     2341 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     2401 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     2461 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     2521 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgagc
     2581 tctagcattt aggtgacact atagaatagg gccctctaga tgcatgctcg agcggccgcc
     2641 aagctttccc ttctggtatc aaaatgctcc ggatcagata tcggcgtagt ccgggacgtc
     2701 gtaggggtag ccgatgtcca cgtcccgcac gtcggtgggt gtgcaagcca ggtccacctc
     2761 ctcctcctcc tcttcctcct cctcggcggc cttggggtcc atgttctgct gggcctggcg
     2821 caggcttgac tccagcagag cttcgatctg ctcctggcag gcccggaggc agtctgggtc
     2881 acacttgatc actctggaga ggaagcgtgt gaggcggtag taggacagga agttgttggg
     2941 gctcctcagg ttcaggcctt gcactgcggc caccacgctc cccgctgcca ccatggaggg
     3001 cggattggaa atgaacttca catctgtggc acagagggca acgaaggtct gcgcgtgttt
     3061 gcggatgatc tgtttgttct cctccgcctc tggcattttg gagaggaagt gttcaatgaa
     3121 atcgtgcggg gtcattgcgg ccaggttcca cttgagcttg ttcaccagga gcagctccat
     3181 ttgcagcagc tcctcgggcc ggatggagtt gtcggtgtag atgcacagct tctcggccgt
     3241 cagggggatg gtctccttca tctcagaggc cacgaacatg caagtggccc ccaggagctg
     3301 caggcggctc tttttcacgg gctccagcga caggaagcgg tccaggtagt tcatggccag
     3361 cgggaagacc tcctcctcgc acttctgttc ctcgcagacc tccagcatcc aggtggcgac
     3421 gatcttccgc atggacggca ggacctcctt ctgcacacat ttgaagtagg acaccgaggg
     3481 cgcgcaggtc tcctccgcct tcagcatggc ccgcagcacc cggtcgttga ggaggttggc
     3541 atcggggtac gcgcggcgga tggtttccac ttcgcagcac aggagctggt gttccatggc
     3601 tggggctctt cctgggcagc tggggagggc tgtgggtcct ggctgggtcc gcgctcggct
     3661 ctcgcttctg ctgccccgcg ctccctcgcg ctcttctgcc cctcgccgga gcgtgcggac
     3721 tctgctgctc gctgctactg cgcccccgaa tttggcggcc gttactagtg gatccgagct
     3781 cggtaccaag cttgggtctc cctatagtga gtcgtattaa tttcgataag ccagtaagca
     3841 gtgggttctc tagttagcca gagagctctg cttatataga cctcccaccg tacacgccta
     3901 ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt
     3961 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     4021 tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg
     4081 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     4141 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     4201 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     4261 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     4321 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     4381 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     4441 caatgtcaac gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta
     4501 agcagattct tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta aattttgctc
     4561 gcgcactact cagcgacctc caacacacaa gcagggagca gatactggct taactatgcg
     4621 gcatcagagc agattgtact gagagtgcac cataggggat cgggagatct cccgatccgt
     4681 cgacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4741 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4801 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4861 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4921 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4981 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5041 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5101 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5161 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5221 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5281 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5341 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5401 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5461 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5521 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5581 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5641 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5821 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5881 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5941 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6001 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6061 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6421 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6481 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6541 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6601 tgagcgagga agcggaag
//
