LOCUS       sequence-64559-e        7371 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 7371)
  TITLE     Ralf Kuehn TALEN plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7371)
  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            "Unpublished"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-TALEN-Rep
FEATURES             Location/Qualifiers
     source          1..7371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(674..690)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gap             1079..1126
                     /estimated_length=48
     primer_bind     complement(1143..1159)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1150..4197
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAELQLSAGRYHYQLVWCQK"
     polyA_signal    4484..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4508..4527)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4562..4581
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4729..4745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4729..4745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4742..4764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4753..4769
                     /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(4777..4783,4784..4801,4802..4807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4822..4843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4960..4977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5211..5230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5890..6681,6682..6750))
                     /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     6513..6532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6751..6855)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6923..6941
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6979..7001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7101..7120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7329..7345
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactcgagg acactgcaga gacctacttc actaacaacc ggtatggtcg
      661 ccagtagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
      721 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
      781 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct
      841 ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg
      901 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
      961 acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt
     1021 gttactcgct cacatttaat gttgatgaaa gctggctata aaaccggtac agttcgaann
     1081 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnntcgc gagtagcttg
     1141 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1201 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1261 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     1321 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     1381 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     1441 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     1501 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     1561 gttaactcgg cgtttcatct gtggtgcaac gggcgctggg tcggttacgg ccaggacagt
     1621 cgtttgccgt ctgaatttga cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg
     1681 gtgatggtgc tgcgctggag tgacggcagt tatctggaag atcaggatat gtggcggatg
     1741 agcggcattt tccgtgacgt ctcgttgctg cataaaccga ctacacaaat cagcgatttc
     1801 catgttgcca ctcgctttaa tgatgatttc agccgcgctg tactggaggc tgaagttcag
     1861 atgtgcggcg agttgcgtga ctacctacgg gtaacagttt ctttatggca gggtgaaacg
     1921 caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta tcgatgagcg tggtggttat
     1981 gccgatcgcg tcacactacg tctgaacgtc gaaaacccga aactgtggag cgccgaaatc
     2041 ccgaatctct atcgtgcggt ggttgaactg cacaccgccg acggcacgct gattgaagca
     2101 gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa atggtctgct gctgctgaac
     2161 ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc atcatcctct gcatggtcag
     2221 gtcatggatg agcagacgat ggtgcaggat atcctgctga tgaagcagaa caactttaac
     2281 gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt acacgctgtg cgaccgctac
     2341 ggcctgtatg tggtggatga agccaatatt gaaacccacg gcatggtgcc aatgaatcgt
     2401 ctgaccgatg atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag
     2461 cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg ggaatgaatc aggccacggc
     2521 gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg cccggtgcag
     2581 tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat gtacgcgcgc
     2641 gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg
     2701 ctacctggag agacgcgccc gctgatcctt tgcgaatacg cccacgcgat gggtaacagt
     2761 cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt atccccgttt acagggcggc
     2821 ttcgtctggg actgggtgga tcagtcgctg attaaatatg atgaaaacgg caacccgtgg
     2881 tcggcttacg gcggtgattt tggcgatacg ccgaacgatc gccagttctg tatgaacggt
     2941 ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg aagcaaaaca ccagcagcag
     3001 tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata cctgttccgt
     3061 catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc gctggcaagc
     3121 ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact gcctgaacta
     3181 ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca accgaacgcg
     3241 accgcatggt cagaagccgg gcacatcagc gcctggcagc agtggcgtct ggcggaaaac
     3301 ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc atctgaccac cagcgaaatg
     3361 gatttttgca tcgagctggg taataagcgt tggcaattta accgccagtc aggctttctt
     3421 tcacagatgt ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc
     3481 cgtgcaccgc tggataacga cattggcgta agtgaagcga cccgcattga ccctaacgcc
     3541 tgggtcgaac gctggaaggc ggcgggccat taccaggccg aagcagcgtt gttgcagtgc
     3601 acggcagata cacttgctga tgcggtgctg attacgaccg ctcacgcgtg gcagcatcag
     3661 gggaaaacct tatttatcag ccggaaaacc taccggattg atggtagtgg tcaaatggcg
     3721 attaccgttg atgttgaagt ggcgagcgat acaccgcatc cggcgcggat tggcctgaac
     3781 tgccagctgg cgcaggtagc agagcgggta aactggctcg gattagggcc gcaagaaaac
     3841 tatcccgacc gccttactgc cgcctgtttt gaccgctggg atctgccatt gtcagacatg
     3901 tataccccgt acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat
     3961 tatggcccac accagtggcg cggcgacttc cagttcaaca tcagccgcta cagtcaacag
     4021 caactgatgg aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac atggctgaat
     4081 atcgacggtt tccatatggg gattggtggc gacgactcct ggagcccgtc agtatcggcg
     4141 gaattacagc tgagcgccgg tcgctaccat taccagttgg tctggtgtca aaaataataa
     4201 taaccgggca ggccatgtct gcccgtattt cgcgtaagga aatccattat gtactattta
     4261 aaaaacacaa acttttggat gttcggttta ttctttttct tttacttttt tatcatggga
     4321 gcctacttcc cgtttttccc gatttggcta catgacatca accatatcag caaaagtgat
     4381 acgggtatta tttttgccgc tatttctctg ttctcgctat tattccaacc gctgtttggt
     4441 ctgctttctg acaaactcgg cctcgactct aggcggccgc ggggatccag acatgataag
     4501 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4561 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4621 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc
     4681 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     4741 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4801 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4861 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6901 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6961 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     7021 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7081 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     7141 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7201 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7261 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7321 acgacgttgt aaaacgacgg ccagtgaatt cgagcttgca tgcctgcagg t
//
