LOCUS       sequence-64564-e        6037 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 6037)
  TITLE     Ralf Kuehn TALEN plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6037)
  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            "Unpublished"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: pCAG-RabZFN-B
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..673
                     /label=chicken beta-actin promoter
     intron          674..1682
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1695..1713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1733..1753
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1763..1786
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2935..2951)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2965..2982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2971..3195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3267..3284)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3267..3283)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3276..3298)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3425..3880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3512..3531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3722..3743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3906..4010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4011..4079,4080..4871)
                     /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(4229..4248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5042..5630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5531..5550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5784..5801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5918..5939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5954..5959,5960..5977,5978..5984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5992..6008
                     /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     5997..6019
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6016..6032
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6016..6032
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccgg attcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
      421 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
      481 attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg
      541 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc gcgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
      841 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
      901 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg cgaggggagc
     1021 gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga ggggaacaaa ggctgcgtgc
     1081 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc ggcggtcggg ctgtaacccc
     1141 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1201 gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1261 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctggcggag ccgaaatctg ggaggcgccg
     1441 ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccatctcca gcctcggggc
     1561 tgccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1681 agatccttaa ttaataatac gactcactat aggggccgcc accatgagat ctcctaagaa
     1741 aaagaggaag gtggcggccg ctgactacaa ggatgacgac gataaaccat ggggtggcgg
     1801 aggtagtcca ttccagtgta gaatctgtat gcgaaacttt tcaacgtctg ggcacttgtc
     1861 caggcacata aggacacaca ccggagagaa acccttcgct tgcgacatat gtggaaggaa
     1921 gttcgcccat aaatggcaga ggaacaagca taccaagatt cacacagggg aaaaaccttt
     1981 ccagtgtcgg atttgtatgc gaaatttctc agaccgctca gttctacgga gacacattcg
     2041 tacccatacc ggcgaaaagc cgtttgcatg cgacatctgt ggcagaaaat ttgctgactc
     2101 cagcactcgc aagaagcaca cgaaaattca caccggacgc cagaaaccct ttcagtgtcg
     2161 aatttgcatg cgtaactttt ctcgatccga ccatctgtcg gaacacatcc gaacccatac
     2221 tggcgaaaag ccattcgcgt gcgacatctg tggtagaaag ttcgccgata agtctaatag
     2281 gaagaagcac acgaaaattc acctgcgcgg atcccagctc gtgaaaagcg aactcgaaga
     2341 aaagaaaagt gaactgcggc acaaactgaa atacgtccca catgaataca ttgagctgat
     2401 cgagattgct aggaactcca cccaggacag aatcctcgag atgaaagtga tggaattctt
     2461 tatgaaagtc tacgggtatc ggggcaagca cctgggcgga tctcgcaaac cagatggggc
     2521 aatctacact gtgggtagtc ccatcgacta tggcgtgatt gtcgatacca aggcctacag
     2581 tgggggttat aatctgccca ttggacaggc tgacgagatg gaacgatacg tggaggaaaa
     2641 ccagacaaga aataagcatc tgaaccccaa tgagtggtgg aaagtgtatc ctagctccgt
     2701 cactgaattc aagtttctct tcgtgtcagg ccactttaag ggaaactaca aagcacagct
     2761 gaccaggctc aatcatatta caaactgcaa tggcgccgtg ctgagcgtcg aggaactgct
     2821 catcggcgga gagatgatca aggccggcac actcaccctg gaggaggtcc gccgaaaatt
     2881 caataacggg gaaatcaact tctgagttta aacgatcacg cgtaaatgat tgcagatcca
     2941 ctagttctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3001 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3061 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3121 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3181 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgaga tccactagtt
     3241 ctagcctcga ggctagagcg gccgccactg gccgtcgttt tacaacgtcg tgactgggaa
     3301 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3361 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3421 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3841 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     3901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4021 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4081 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4141 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4201 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4261 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4321 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4381 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4441 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4501 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4561 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4621 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4681 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4741 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4801 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4861 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4921 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4981 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5161 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5701 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5761 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5821 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5881 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     5941 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     6001 gataacaatt tcacacagga aacagctatg accatga
//
