LOCUS       sequence-127179-        7432 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 7432)
  TITLE     Li-Huei Tsai's Plasmids.
REFERENCE   2  (bases 1 to 7432)
  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"
COMMENT     Sequence Label: pCMV-P35
FEATURES             Location/Qualifiers
     source          1..7432
                     /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"
     primer_bind     561..580
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    579..634
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     680..699
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          1785..2357
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2448..2472)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2469..2672)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2498..2518)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2673..3052
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3345..3392
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3417..3436
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3630..4424)
                     /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(3741..3760)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4351..4370
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4481..4626)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(5037..5054)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5037..5053)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5046..5068)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5262..5281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5381..5403
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5441..5459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5527..5631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5632..5700,5701..6492)
                     /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(5850..5869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6663..7251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7152..7171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7405..7422
                     /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 catgattacg aattgcggcc
      241 gatccgtcga gggatctcca taagagaaga gggacagcta tgactgggag tagtcaggag
      301 aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa
      361 aataacacaa aacaaaatat aaaaaaaatc taacctcaag tcaaggcttt tctatggaat
      421 aaggaatgga cagcaggggg ctgtttcata tactgatgac ctctttatag ccaacctttg
      481 ttcatggcag ccagcatatg ggcatatgtt gccaaactct aaaccaaata ctcattctga
      541 tgttttaaat gatttgccct cccatatgtc cttccgagtg agagacacaa aaaattccaa
      601 cacactattg caatgaaaat aaatttcctt tattagccag aagtcagatg ctcaaggggc
      661 ttcatgatgt ccccataatt tttggcagag ggaaaaagat ctcagtggta tttgtgagcc
      721 agggcattgg ccacaccagc caccaccttc tgataggcag cctgcacctg aggagtgaat
      781 tcctcgacgg atcctcaccg atccaggcct aggaggagcc gcttcttgtc ctcctggccg
      841 ctctcgttct tcaggtcgga gaagacctgt gtgaagtagt gtgggtcggc atttatctgc
      901 agcatctttg agctcatgag gttgatgaca gagaggcaac ggtcccaaaa ggcctccttg
      961 cagctctcca ccaggaaggg cttgagcggg taggagatct cgttgcccat gtaggagtag
     1021 gagaggtaca ggcatgtcag caggacggcc tggagctcgt gatccgagcc cacctcggag
     1081 gagataacat ccctgcagag catgtagagg aagaccacgt tggccggcgt gatgaagccc
     1141 tggtcctgcc agccctgcag aagcagcgag cggtccacgc tgcgcagcca gagcacgggg
     1201 tccgtggggg acaggtgctt caggcggtag caccggcggc agagaaactc acccaggcag
     1261 cgaagcagct cactggtgga cgcctggacg atgacccgtt tgggcgtccc tgcggaggtg
     1321 acggcagggt gaggggcttt cttgactgag gaggagcccc cggtctggga acccgagagc
     1381 tggctggccg ggggtgcagg cggctgggcc ggtgggggct gggcgaatgt ggacaggttg
     1441 gcgcacgaca gcgacttctt caggttctca ttgttgaggt gcgtgatgtt gttctggtag
     1501 ctgctgttag gctgcacctt cttggagttc ttcttcttgg ccgacacggc cacgattctc
     1561 ttccaaggca gcacggagat gatggagtgg cgcttcaggt tcttgtcctt ggcgttcttg
     1621 ctgttctgta cggccgtata gtggcccacg gtggccgcgc catcctcaaa cagcgtggcc
     1681 ttccggtagc tgggagacag ggacagcacc gtgcccatgg atccgtcgag gaattctttg
     1741 ccaaaatgat gagacagcac aacaaccagc acgttgccca ggagctgtag gaaagagaag
     1801 aaggcatgaa catggttagc agaggggccc ggtttggact cagagtattt tatcctcatc
     1861 tcaaacagtg tatatcattg taaccataaa gagaaaggca ggatgatgac cagggtgtag
     1921 ttgtttctac caataagaat atttccacgc cagccagaat ttatatgcag aaatattcta
     1981 ccttatcatt taattataac aattgttctc taaaactgtg ctgaagtaca atataatata
     2041 ccctgattgc cttgaaaaaa aagtgattag agaaagtact tacaatctga caaataaaca
     2101 aaagtgaatt taaaaattcg ttacaaatgc aagctaaagt ttaacgaaaa agttacagaa
     2161 aatgaaaaga aaataagagg agacaatggt tgtcaacaga gtagaaagtg aaagaaacaa
     2221 aattatcatg agggtccatg gtgatacaag ggacatcttc ccattctaaa caacaccctg
     2281 aaaactttgc cccctccata taacatgaat tttacaatag cgaaaaagaa agaacaatca
     2341 agggtcccca aactcaccct gaagttctca ggatcggtcg accggaggct ggatcggtcc
     2401 cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc
     2461 actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2521 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     2581 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     2641 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     2701 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     2761 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     2821 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     2881 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     2941 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     3001 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcaacatggc
     3061 ggtaatgttg gacatgagcc aatataaatg tacatattat gatatggata caacgtatgc
     3121 aatgggccaa gctctctaga gggccgcgat ccgccgtccc taacccacgg ggcccgtggc
     3181 tatggcaggg cttgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg
     3241 ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg
     3301 ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac
     3361 accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt
     3421 ctccttccgt gtttcagtta gcctccccca tctcccgggg tgggcgaaga actccagcat
     3481 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     3541 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
     3601 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     3661 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     3721 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     3781 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     3841 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     3901 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     3961 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     4021 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     4081 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     4141 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     4201 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     4261 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     4321 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     4381 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     4441 cctgtctctt gatcagatct gcggcacgct gttgacgctg ttaagcgggt cgctgcaggg
     4501 tcgctcggtg ttcgaggcca cacgcgtcac cttaatatgc gaagtggacc tgggaccgcg
     4561 ccgccccgac tgcatctgcg tgttcgaatt cgccaatgac aagacgctgg gcggggtttg
     4621 tgtcatcata gaactaaaga catgcaaata tatttcttcc ggggacaccg ccagcaaacg
     4681 cgagcaacgg gccacgggga tgaagcagga tcttgatccc ctgcgccatc agatccttgg
     4741 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     4801 agctggcaat tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt
     4861 aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc
     4921 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtgtt
     4981 ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg cagcgtgaag cttggcactg
     5041 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5101 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5161 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     5221 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     5281 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5341 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5401 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5461 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5521 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5581 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5641 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5701 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5761 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5821 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5881 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5941 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6001 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6061 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6121 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6181 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6241 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6301 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6361 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6421 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6481 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6541 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6721 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6781 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6841 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6901 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6961 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7021 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7081 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7141 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7201 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7261 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7321 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7381 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
