LOCUS       sequence-382117-        7592 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 7592)
  AUTHORS   Kurosu T, Zhang F, Peterlin BM
  TITLE     Transcriptional activity and substrate recognition of cyclin T2 from
            P-TEFb.
  JOURNAL   Gene. 2004 Dec 8. 343(1):173-9.
  PUBMED    15563843
REFERENCE   2  (bases 1 to 7592)
  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: "Gene. 2004 
            Dec 8. 343(1):173-9."
COMMENT     Sequence Label: pCMV Flag cycT2b (P#1718)
FEATURES             Location/Qualifiers
     source          1..7592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             670..693
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2911..2929)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2912..2929)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2949..2966)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2955..3179
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3225..3653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3312..3331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3522..3543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3662..3682)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3667..3996
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3847..3982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3909..3928
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4063..4857
                     /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(4117..4136)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4727..4746
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5031..5152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5068..5087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5122..5141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5201..5217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5201..5217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5214..5236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5225..5241
                     /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(5249..5255,5256..5273,5274..5279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5294..5315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5432..5449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5603..6191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5683..5702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6362..7153,7154..7222))
                     /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     6985..7004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7223..7327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7402..7421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccatgg attacaagga cgacgatgac aaggaattcc gtcgacggat ggcgtcgggc
      721 cgtggagctt cttctcgctg gttctttact cgggaacagc tggagaacac gccgagccgc
      781 cgctgcggag tggaggcgga taaagagctc tcgtgccgcc agcaggcggc caacctcatc
      841 caggagatgg gacagcgtct caatgtctct cagcttacaa taaacactgc gattgtttat
      901 atgcacaggt tttatatgca ccattctttc accaaattca acaaaaatat aatatcgtct
      961 actgcattat ttttggctgc aaaagtggaa gaacaggctc gaaaacttga acatgttatc
     1021 aaagtagcac atgcttgtct tcatcctcta gagccactgc tggatactaa atgtgatgct
     1081 taccttcaac agactcaaga actggttata cttgaagcca taatgctaca aactctaggt
     1141 tttgagatca ccattgaaca cccacacaca gatgtggtga aatgtaccca gttagtaaga
     1201 gcaagcaagg atttggcaca gacatcctat ttcatggcta ccaacagtct gcatcttaca
     1261 accttctgtc ttcagtacaa accaacagtg atagcatgtg tatgcattca tttggcttgc
     1321 aaatggtcca attgggagat ccctgtatca actgatggaa agcattggtg ggaatatgtg
     1381 gatcctacag ttactctaga attattagat gagctaacac atgagtttct acaaatattg
     1441 gagaaaacgc ctaataggtt gaagaagatt cgaaactgga gggctaatca ggcagctagg
     1501 aaaccaaaag tagatggaca ggtatcagag acaccacttc ttggttcatc tttggtccag
     1561 aattccattt tagtagatag tgtcactggt gtgcctacaa acccaagttt tcagaaacca
     1621 tctacatcag cattccctgc gccagtacct ctaaattcag gaaatatttc tgttcaagac
     1681 agccatacat ctgataattt gtcaatgcta gcaacaggaa tgccaagtac ttcatacggt
     1741 ttatcatcac accaggaatg gcctcaacat caagactcag caaggacaga acagctatat
     1801 tcacagaaac aggagacatc tttgtctggt agccagtaca acatcaactt ccagcaggga
     1861 ccttctatat cactgcattc aggattacat cacagacctg acaaaatttc agatcattct
     1921 tctgttaagc aagaatatac tcataaagca gggagcagta aacaccatgg gccaatttcc
     1981 actactccag gaataattcc tcagaaaatg tctttagata aatatagaga aaagcgtaaa
     2041 ctagaaactc ttgatctcga tgtaagggat cattatatag ctgcccaggt agaacagcag
     2101 cacaaacaag ggcagtcaca ggcagccagc agcagttctg ttacttctcc cattaaaatg
     2161 aaaataccta tcgcaaatac tgaaaaatac atggcagata aaaaggaaaa gagtgggtca
     2221 ctgaaattac ggattccaat accacccact gataaaagcg ccagtaaaga agaactgaaa
     2281 atgaaaataa aagtttcttc ttcagaaaga cacagctctt ctgatgaagg cagtgggaaa
     2341 agcaaacatt caagcccaca tattagcaga gaccataagg agaagcacaa ggagcatcct
     2401 tcaagccgcc accacaccag cagccacaag cattcccact cgcatagtgg cagcagcagc
     2461 ggtggcagta aacacagtgc cgacggaata ccacccactg ttctgaggag tcctgttggc
     2521 ctgagcagtg atggcatttc ctctagctcc agctcttcaa ggaagaggct gcatgtcaat
     2581 gatgcatctc acaaccacca ctccaaaatg agcaaaagtt ccaaaagttc aggtagttca
     2641 tctagttctt cctcctctgt taagcagtat atatcctctc acaactctgt ttttaaccat
     2701 cccttacccc ctcctccccc tgtcacatac caggtgggct acggacatct cagcaccctc
     2761 gtgaaactgg acaagaagcc agtggagacc aacggtcctg atgccaatca cgagtacagt
     2821 acaagcagcc agcatatgga ctacaaagac acattcgaca tgctggactc actgttaagt
     2881 gcccaaggaa tgaacatgta agggccctat tctatagtgt cacctaaatg ctagagctcg
     2941 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3001 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3061 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3121 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3181 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3661 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3721 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3901 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3961 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4021 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4081 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4141 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4201 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4261 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4321 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4381 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4441 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4501 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4561 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4621 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4681 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4741 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4801 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4861 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4921 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4981 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5041 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5101 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5161 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5221 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5281 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5341 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5401 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5461 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5521 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6121 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6301 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6961 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7381 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     7441 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     7501 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     7561 gcttcgcgat gtacgggcca gatatacgcg tt
//
