LOCUS       sequence-64565-e        5566 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 5566)
  TITLE     Ralf Kuehn TALEN plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5566)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCMV-Luc
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     700..724
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      830..839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             836..2488
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(902..920)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2529..2650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2540..2559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2594..2613
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2898..2915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3069..3657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3149..3168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3828..4619,4620..4688))
                     /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     4451..4470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4689..4793)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4820..5275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4907..4926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5117..5138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5406..5454
                     /note="synthetic polyadenylation signal"
     misc_feature    5468..5559
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5508..5527
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgaggag cttggcccat tgcatacgtt
       61 gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg
      121 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acgctaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
      721 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      781 cagcctccgc ggccccgaat tagcttggca ttccggtact gttggtaaag ccaccatgga
      841 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
      901 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
      961 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
     1021 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     1081 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     1141 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
     1201 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     1261 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
     1321 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     1381 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
     1441 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
     1501 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     1561 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     1621 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
     1681 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
     1741 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     1801 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     1861 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     1921 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     1981 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     2041 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     2101 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     2161 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     2221 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     2281 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     2341 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     2401 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     2461 caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc
     2521 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2581 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2641 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2701 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     2761 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     2821 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     2881 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2941 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3241 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3481 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3841 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3901 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3961 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4021 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4081 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4141 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4201 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4261 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4321 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4381 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4441 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4501 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4561 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4621 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4681 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4741 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4801 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4861 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4921 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4981 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5041 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5101 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5161 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5221 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     5281 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5341 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     5401 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     5461 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     5521 ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata
//
