LOCUS       sequence-74785-e        6339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEYellow-Telomere for In Vitro Translation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6339)
  AUTHORS   Ma H, Reyes-Gutierrez P, Pederson T
  TITLE     Visualization of repetitive DNA sequences in human chromosomes with 
            transcription activator-like effectors.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Dec 9.
  PUBMED    24324157
REFERENCE   2  (bases 1 to 6339)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Dec 9."
COMMENT     Sequence Label: pT7CFE1-TelR15-YPet
FEATURES             Location/Qualifiers
     source          1..6339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    70..531
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(126..143)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(2582..2584,2585..2587,2588..3301)
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     primer_bind     complement(2888..2907)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3323..3340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3398..3416)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3412..3459
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3488..3505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3659..4247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3739..3758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4418..5209,5210..5278))
                     /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     5041..5060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5279..5383)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5451..5469
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5507..5529)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5629..5648
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5717..6172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5854..5875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6066..6085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6302..6324
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6316..6333
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6317..6333
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattaatt ccggttattt tccaccatat tgccgtcttt
       61 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
      121 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
      181 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
      241 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
      301 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctcac
      361 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
      421 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta
      481 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
      541 cacccatatg ggatccggta tccacggagt cccagcagcc gtagatttga gaactttggg
      601 atattcacag cagcagcagg aaaagatcaa gcccaaagtg aggtcgacag tcgcgcagca
      661 tcacgaagcg ctggtgggtc atgggtttac acatgcccac atcgtagcct tgtcgcagca
      721 ccctgcagcc cttggcacgg tcgccgtcaa gtaccaggac atgattgcgg cgttgccgga
      781 agccacacat gaggcgatcg tcggtgtggg gaaacagtgg agcggagccc gagcgcttga
      841 ggccctgttg acggtcgcgg gagagctgag agggcctccc cttcagctgg acacgggcca
      901 gttgctgaag atcgcgaagc ggggaggagt cacggcggtc gaggcggtgc acgcgtggcg
      961 caatgcgctc acgggagcac ccctcaacct gaccccagag caggtcgtgg caattgcgag
     1021 caacatcggg ggaaagcagg cactcgaaac cgtccagagg ttgctgcctg tgctgtgcca
     1081 agcgcacgga cttacgccag agcaggtcgt ggcaattgcg agcaacatcg ggggaaagca
     1141 ggcactcgaa accgtccaga ggttgctgcc tgtgctgtgc caagcgcacg gactaacccc
     1201 agagcaggtc gtggcaattg cgagccatga cgggggaaag caggcactcg aaaccgtcca
     1261 gaggttgctg cctgtgctgt gccaagcgca cgggttgacc ccagagcagg tcgtggcaat
     1321 tgcgagccat gacgggggaa agcaggcact cgaaaccgtc cagaggttgc tgcctgtgct
     1381 gtgccaagcg cacggcctga ccccagagca ggtcgtggca attgcgagcc atgacggggg
     1441 aaagcaggca ctcgaaaccg tccagaggtt gctgcctgtg ctgtgccaag cgcacggact
     1501 gacaccagag caggtcgtgg caattgcgag caacggaggg ggaaagcagg cactcgaaac
     1561 cgtccagagg ttgctgcctg tgctgtgcca agcgcacgga cttacacccg aacaagtcgt
     1621 ggcaattgcg agcaacatcg ggggaaagca ggcactcgaa accgtccaga ggttgctgcc
     1681 tgtgctgtgc caagcgcacg gacttacgcc agagcaggtc gtggcaattg cgagcaacat
     1741 cgggggaaag caggcactcg aaaccgtcca gaggttgctg cctgtgctgt gccaagcgca
     1801 cggactaacc ccagagcagg tcgtggcaat tgcgagccat gacgggggaa agcaggcact
     1861 cgaaaccgtc cagaggttgc tgcctgtgct gtgccaagcg cacgggttga ccccagagca
     1921 ggtcgtggca attgcgagcc atgacggggg aaagcaggca ctcgaaaccg tccagaggtt
     1981 gctgcctgtg ctgtgccaag cgcacggcct gaccccagag caggtcgtgg caattgcgag
     2041 ccatgacggg ggaaagcagg cactcgaaac cgtccagagg ttgctgcctg tgctgtgcca
     2101 agcgcacgga ctgacaccag agcaggtcgt ggcaattgcg agcaacggag ggggaaagca
     2161 ggcactcgaa accgtccaga ggttgctgcc tgtgctgtgc caagcgcacg gcctcacccc
     2221 agagcaggtc gtggcaattg cgagcaacat cgggggaaag caggcactcg aaaccgtcca
     2281 gaggttgctg cctgtgctgt gccaagcgca cggactcacg cctgagcagg tagtggctat
     2341 tgcatccaac atcgggggca gacccgcact ggagtcaatc gtggcccagc tttcgaggcc
     2401 ggaccccgcg ctggccgcac tcactaatga tcatcttgta gcgctggcct gcctcggcgg
     2461 acgacccgcc ttggatgcgg tgaagaaggg gctcccgcac gcgcctgcat tgattaagcg
     2521 gaccaacaga aggattcccg agaggacatc acatcgagtg gcaggcctgc agggaagtgg
     2581 aatggtgagc aaaggcgaag agctgttcac cggcgtggtg cccatcctgg tggagctgga
     2641 cggcgacgtg aacggccaca agttcagcgt gagcggcgag ggcgagggcg acgccaccta
     2701 cggcaagctg accctgaagc tgctgtgcac caccggcaag ctgcccgtgc cctggcccac
     2761 cctggtgacc accctgggct acggcgtgca gtgcttcgcc cggtaccccg accacatgaa
     2821 gcagcacgac ttcttcaaga gcgccatgcc cgagggctac gtgcaggagc ggaccatctt
     2881 cttcaaggac gacggcaact acaagacccg ggccgaggtg aagttcgagg gcgacaccct
     2941 ggtgaaccgg atcgagctga agggcatcga cttcaaggag gacggcaaca tcctgggcca
     3001 caagctggag tacaactaca acagccacaa cgtgtacatc accgccgaca agcagaagaa
     3061 cggcatcaag gccaacttca agatccggca caacatcgag gacggcggcg tgcagctggc
     3121 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3181 ctacctgagc taccagagcg ccctgttcaa ggaccccaac gagaagcggg accacatggt
     3241 gctgctggag ttcctgaccg ccgccggcat caccgagggc atgaacgagc tctataagta
     3301 gtctagagcg gccgcactcg agcaccacca ccaccaccac tgagatctga ctgaaaaaaa
     3361 aaaaaaaaaa aaaaaaaaaa aaagtttaaa cactagtccg ctgagcaata actagcataa
     3421 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3481 gggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3541 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3601 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3661 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3721 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3781 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3841 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3901 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3961 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4021 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4081 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4141 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4201 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4261 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4321 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4381 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4441 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4501 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4561 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4621 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4681 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4741 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4801 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4861 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4921 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4981 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5041 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5101 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5161 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5221 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5281 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5341 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5401 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5461 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     5521 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     5581 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     5641 agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5701 aaaataccgc atcaggaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     5761 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5821 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5881 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5941 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     6001 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     6061 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     6121 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcgcgccat
     6181 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6241 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6301 tcccagtcac gacgttgtaa aacgacggcc agtgaattg
//
