LOCUS       sequence-222662-        7455 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 7455)
  AUTHORS   Ballard DW, Dixon EP, Peffer NJ, Bogerd H, Doerre S, Stein B, Greene
            WC
  TITLE     The 65-kDa subunit of human NF-kappa B functions as a potent 
            transcriptional activator and a target for v-Rel-mediated 
            repression.
  JOURNAL   Proc Natl Acad Sci U S A. 1992 Mar 1. 89(5):1875-9.
  PUBMED    1542686
REFERENCE   2  (bases 1 to 7455)
  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. 1992 Mar 1. 89(5):1875-9."
COMMENT     Sequence Label: pCMV4 p65
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     738..754
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     738..754
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2131..2502
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     polyA_signal    3347..3969
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3993..4013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3998..4327
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4178..4313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4366..4462
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(4538..4557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4539..4557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4571..4587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4571..4587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4584..4606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4595..4611
                     /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(4619..4625,4626..4643,4644..4649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4664..4685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4802..4819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4973..5561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5053..5072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5732..6523,6524..6592))
                     /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     6355..6374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6593..6697)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6724..7179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6811..6830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7021..7042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7305..7327
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7319..7336
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7320..7336
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattgtt tttattttta
      661 attttctttc aaatacttcc atcgaattca gatctggtac cacgcgtatc gataagcttg
      721 catgcctgca ggtcgactct agaactagtg gatcccccgg gctgcaggaa ttccggagtt
      781 ccatggctcg tctgtagtgc acgccgcggg cccagctgcg accccggccc cgcccccggg
      841 accccggcca tggacgaact gttccccctc atcttcccgg cagagccagc ccaggcctct
      901 ggcccctatg tggagatcat tgagcagccc aagcagcggg gcatgcgctt ccgctacaag
      961 tgcgaggggc gctccgcggg cagcatccca ggcgagagga gcacagatac caccaagacc
     1021 caccccacca tcaagatcaa tggctacaca ggaccaggga cagtgcgcat ctccctggtc
     1081 accaaggacc ctcctcaccg gcctcacccc cacgagcttg taggaaagga ctgccgggat
     1141 ggcttctatg aggctgagct ctgcccggac cgctgcatcc acagtttcca gaacctggga
     1201 atccagtgtg tgaagaagcg ggacctggag caggctatca gtcagcgcat ccagaccaac
     1261 aacaacccct tccaagttcc tatagaagag cagcgtgggg actacgacct gaatgctgtg
     1321 cggctctgct tccaggtgac agtgcgggac ccatcaggca ggcccctccg cctgccgcct
     1381 gtcctttctc atcccatctt tgacaatcgt gcccccaaca ctgccgagct caagatctgc
     1441 cgagtgaacc gaaactctgg cagctgcctc ggtggggatg agatcttcct actgtgtgac
     1501 aaggtgcaga aagaggacat tgaggtgtat ttcacgggac caggctggga ggcccgaggc
     1561 tccttttcgc aagctgatgt gcaccgacaa gtggccattg tgttccggac ccctccctac
     1621 gcagacccca gcctgcaggc tcctgtgcgt gtctccatgc agctgcggcg gccttccgac
     1681 cgggagctca gtgagcccat ggaattccag tacctgccag atacagacga tcgtcaccgg
     1741 attgaggaga aacgtaaaag gacatatgag accttcaaga gcatcatgaa gaagagtcct
     1801 ttcagcggac ccaccgaccc ccggcctcca cctcgacgca ttgctgtgcc ttcccgcagc
     1861 tcagcttctg tccccaagcc agcaccccag ccctatccct ttacgtcatc cctgagcacc
     1921 atcaactatg atgagtttcc caccatggtg tttccttctg ggcagatcag ccaggcctcg
     1981 gccttggccc cggcccctcc ccaagtcctg ccccaggctc cagcccctgc ccctgctcca
     2041 gccatggtat cagctctggc ccaggcccca gcccctgtcc cagtcctagc cccaggccct
     2101 cctcaggctg tggccccacc tgcccccaag cccacccagg ctggggaagg aacgctgtca
     2161 gaggccctgc tgcagctgca gtttgatgat gaagacctgg gggccttgct tggcaacagc
     2221 acagacccag ctgtgttcac agacctggca tccgtcgaca actccgagtt tcagcagctg
     2281 ctgaaccagg gcatacctgt ggccccccac acaactgagc ccatgctgat ggagtaccct
     2341 gaggctataa ctcgcctagt gacaggggcc cagaggcccc ccgacccagc tcctgctcca
     2401 ctgggggccc cggggctccc caatggcctc ctttcaggag atgaagactt ctcctccatt
     2461 gcggacatgg acttctcagc cctgctgagt cagatcagct cctaaggggg tgacgcctgc
     2521 cctccccaga gcactgggtt gcaggggatt gaagccctcc aaaagcactt acggattctg
     2581 gtggggtgtg ttccaactgc ccccaacttt gtggatgtct tccttggagg ggggagccat
     2641 attttattct tttattgtca gtatctgtat ctctctctct ttttggaggt gcttaagcag
     2701 aagcattaac ttctctggaa aggggggagc tggggaaact caaacttttc ccctgtcctg
     2761 atggtcagct cccttctctg tagggaactc tggggtcccc catccccatc ctccagcttc
     2821 tggtactctc ctagagacag aagcaggctg gaggtaaggc ctttgagccc acaaagcctt
     2881 atcaagtgtc ttccatcatg gattcattac agcttaatca aaataacgcc ccagatacca
     2941 gcccctgtat ggcactggca ttgtccctgt gcctaacacc agcgtttgag gggctggcct
     3001 tcctgcccta cagaggtctc tgccggctct ttccttgctc aaccatggct gaaggaaacc
     3061 agtgcaacag cactggctct ctccaggatc cagaaggggt ttggtctggg acttccttgc
     3121 tctccctctt ctcaagtgcc ttaatagtag ggtaagttgt taagagtggg ggagagcagg
     3181 ctggcagctc tccagtcagg aggcatagtt tttactgaac aatcaaagca cttggactct
     3241 tgctctttct actctgaact aataaatctg ttgccaagct gaaaaaaaaa aaaaaaggaa
     3301 ttcgatatca agcttgcatg cctgcaggtc gactctagag gatcccgggt ggcatccctg
     3361 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3421 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3481 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3541 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3601 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3661 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3721 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3781 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3841 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3901 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3961 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     4021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4081 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4141 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4201 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4261 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4321 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
     4381 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
     4441 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgct
     4501 gcaggtcgac tctagggccg cgtaccgagc tcgaattccc tatagtgagt cgtattaaat
     4561 tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4621 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4681 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4741 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4801 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5401 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5701 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5761 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5821 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5881 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5941 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6001 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6061 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6121 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6181 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6241 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6301 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6361 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6421 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6481 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6541 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6601 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6661 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6721 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6781 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6841 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6901 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6961 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     7021 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     7081 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     7141 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     7201 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     7261 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     7321 taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc aattagccat
     7381 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     7441 tctatatcat aatat
//
