LOCUS       sequence-74782-e        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEBlue-Telomere for In Vitro Translation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  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 6333)
  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-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..6333
                     /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..3292
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             3317..3334
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3392..3410)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3406..3453
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3482..3499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3653..4241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3733..3752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4412..5203,5204..5272))
                     /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     5035..5054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5273..5377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5445..5463
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5501..5523)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5623..5642
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5711..6166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5848..5869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6060..6079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6296..6318
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6310..6327
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6311..6327
                     /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 aatggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga
     2641 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     2701 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     2761 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     2821 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     2881 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     2941 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     3001 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     3061 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acgccaagac
     3121 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     3181 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     3241 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attagtctag
     3301 agcggccgca ctcgagcacc accaccacca ccactgagat ctgactgaaa aaaaaaaaaa
     3361 aaaaaaaaaa aaaaaaagtt taaacactag tccgctgagc aataactagc ataacccctt
     3421 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccgggctt
     3481 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3541 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4081 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4261 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4321 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4381 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4441 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4501 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4561 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4621 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4681 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4741 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4801 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4861 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4921 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4981 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5041 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5101 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5161 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5221 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5281 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5341 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5401 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5461 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5521 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5581 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     5641 tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5701 ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5761 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5821 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5881 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5941 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6001 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6061 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6121 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg ccattcgcca
     6181 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6241 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6301 tcacgacgtt gtaaaacgac ggccagtgaa ttg
//
