LOCUS       sequence-74784-e        6336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEGreen-Telomere for In Vitro Translation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  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 6336)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..6336
                     /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             2582..3295
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3320..3337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3395..3413)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3409..3456
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3485..3502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3656..4244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3736..3755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4415..5206,5207..5275))
                     /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     5038..5057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5276..5380)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5448..5466
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5504..5526)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5626..5645
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5714..6169)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5851..5872)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6063..6082
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6299..6321
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6313..6330
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6314..6330
                     /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 aatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
     2641 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
     2701 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt ggccgactct
     2761 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
     2821 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
     2881 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
     2941 aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc tgggccataa
     3001 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
     3061 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
     3121 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
     3181 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
     3241 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatagtc
     3301 tagagcggcc gcactcgagc accaccacca ccaccactga gatctgactg aaaaaaaaaa
     3361 aaaaaaaaaa aaaaaaaaaa gtttaaacac tagtccgctg agcaataact agcataaccc
     3421 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccggg
     3481 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3541 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3661 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4081 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4321 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4381 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4441 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4501 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4561 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4621 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4681 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4741 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4801 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4861 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4921 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4981 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5041 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5101 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5161 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5221 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5281 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5341 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5401 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5461 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5521 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5581 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     5641 ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5701 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     5761 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5821 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5881 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5941 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6001 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6061 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6121 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     6181 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6241 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     6301 cagtcacgac gttgtaaaac gacggccagt gaattg
//
