LOCUS       sequence-74786-e        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALERed-Telomere for In Vitro Translation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  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 6330)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..6330
                     /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="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2727..2745)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2995..3014
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3314..3331
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3389..3407)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3403..3450
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3479..3496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3650..4238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3730..3749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4409..5200,5201..5269))
                     /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     5032..5051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5270..5374)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5442..5460
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5498..5520)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5620..5639
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5708..6163)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5845..5866)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6057..6076
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6293..6315
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6307..6324
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6308..6324
                     /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 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     2641 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2701 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2761 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     2821 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2881 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct cactgcagga
     2941 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     3001 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     3061 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     3121 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     3181 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3241 acgcgctgag ggccgccact ccaccggcgg catggacgag ctgtacaagt agtctagagc
     3301 ggccgcactc gagcaccacc accaccacca ctgagatctg actgaaaaaa aaaaaaaaaa
     3361 aaaaaaaaaa aaaagtttaa acactagtcc gctgagcaat aactagcata accccttggg
     3421 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cgggcttcct
     3481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4081 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4381 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5401 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5461 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     5521 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     5581 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     5641 tgagagtgca ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5701 catcaggaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5761 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5821 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5881 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5941 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6001 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6061 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6121 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcgcgcca ttcgccattc
     6181 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6241 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6301 cgacgttgta aaacgacggc cagtgaattg
//
