LOCUS       sequence-64561-e        7386 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 7386)
  TITLE     Ralf Kuehn TALEN plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7386)
  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            "Unpublished"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-Rab-Rep
FEATURES             Location/Qualifiers
     source          1..7386
                     /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"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(680..696)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1082..1091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1158..1174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1165..4212
                     /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    4499..4633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4523..4542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4577..4596
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4744..4760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4744..4760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4757..4779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4768..4784
                     /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(4792..4798,4799..4816,4817..4822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4837..4858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4975..4992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5146..5734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5226..5245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5905..6696,6697..6765))
                     /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     6528..6547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6766..6870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6938..6956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6994..7016)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7116..7135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7344..7360
                     /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 gtactcgacg acactgcaga gacctacttc actaacaacc ggtgccacca
      661 tggtcgcgag tagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
      721 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
      781 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct
      841 ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg
      901 aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct
      961 acaccaacgt gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga
     1021 cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctataaacc ggtacagttc
     1081 ggccaccatg gtcgctatgt gcaccaaaac ttctcctcgc actaccgggc caccattggt
     1141 gtggcgagta gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1201 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1261 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt
     1321 gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag
     1381 gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac
     1441 accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg
     1501 ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga
     1561 attatttttg atggcgttaa ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt
     1621 tacggccagg acagtcgttt gccgtctgaa tttgacctga gcgcattttt acgcgccgga
     1681 gaaaaccgcc tcgcggtgat ggtgctgcgc tggagtgacg gcagttatct ggaagatcag
     1741 gatatgtggc ggatgagcgg cattttccgt gacgtctcgt tgctgcataa accgactaca
     1801 caaatcagcg atttccatgt tgccactcgc tttaatgatg atttcagccg cgctgtactg
     1861 gaggctgaag ttcagatgtg cggcgagttg cgtgactacc tacgggtaac agtttcttta
     1921 tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga aattatcgat
     1981 gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa cccgaaactg
     2041 tggagcgccg aaatcccgaa tctctatcgt gcggtggttg aactgcacac cgccgacggc
     2101 acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat tgaaaatggt
     2161 ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca cgagcatcat
     2221 cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatatcct gctgatgaag
     2281 cagaacaact ttaacgccgt gcgctgttcg cattatccga accatccgct gtggtacacg
     2341 ctgtgcgacc gctacggcct gtatgtggtg gatgaagcca atattgaaac ccacggcatg
     2401 gtgccaatga atcgtctgac cgatgatccg cgctggctac cggcgatgag cgaacgcgta
     2461 acgcgaatgg tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc gctggggaat
     2521 gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct
     2581 tcccgcccgg tgcagtatga aggcggcgga gccgacacca cggccaccga tattatttgc
     2641 ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa atggtccatc
     2701 aaaaaatggc tttcgctacc tggagagacg cgcccgctga tcctttgcga atacgcccac
     2761 gcgatgggta acagtcttgg cggtttcgct aaatactggc aggcgtttcg tcagtatccc
     2821 cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa atatgatgaa
     2881 aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa cgatcgccag
     2941 ttctgtatga acggtctggt ctttgccgac cgcacgccgc atccagcgct gacggaagca
     3001 aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc
     3061 gaatacctgt tccgtcatag cgataacgag ctcctgcact ggatggtggc gctggatggt
     3121 aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt
     3181 gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta
     3241 gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca tcagcgcctg gcagcagtgg
     3301 cgtctggcgg aaaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg
     3361 accaccagcg aaatggattt ttgcatcgag ctgggtaata agcgttggca atttaaccgc
     3421 cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg
     3481 cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc
     3541 attgacccta acgcctgggt cgaacgctgg aaggcggcgg gccattacca ggccgaagca
     3601 gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac gaccgctcac
     3661 gcgtggcagc atcaggggaa aaccttattt atcagccgga aaacctaccg gattgatggt
     3721 agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg
     3781 cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta
     3841 gggccgcaag aaaactatcc cgaccgcctt actgccgcct gttttgaccg ctgggatctg
     3901 ccattgtcag acatgtatac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg
     3961 acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt caacatcagc
     4021 cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa
     4081 ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc
     4141 ccgtcagtat cggcggaatt acagctgagc gccggtcgct accattacca gttggtctgg
     4201 tgtcaaaaat aataataacc gggcaggcca tgtctgcccg tatttcgcgt aaggaaatcc
     4261 attatgtact atttaaaaaa cacaaacttt tggatgttcg gtttattctt tttcttttac
     4321 ttttttatca tgggagccta cttcccgttt ttcccgattt ggctacatga catcaaccat
     4381 atcagcaaaa gtgatacggg tattattttt gccgctattt ctctgttctc gctattattc
     4441 caaccgctgt ttggtctgct ttctgacaaa ctcggcctcg actctaggcg gccgcgggga
     4501 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4561 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4621 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4681 gtgggaggtt ttttcggatc ctctagagtc gacctgcagg catgcaagct tggcgtaatc
     4741 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4801 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4861 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5581 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6901 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6961 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     7021 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7081 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7141 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     7201 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7261 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7321 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc ttgcatgcct
     7381 gcaggt
//
