LOCUS       sequence-322239-        5745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Murine Leukemia Reverse Transcriptase (MLRT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5745)
  AUTHORS   Elbaz J, Yin P, Voigt CA
  TITLE     Genetic encoding of DNA nanostructures and their self-assembly in 
            living bacteria.
  JOURNAL   Nat Commun. 2016 Apr 19;7:11179. doi: 10.1038/ncomms11179.
  PUBMED    27091073
REFERENCE   2  (bases 1 to 5745)
  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: "Nat 
            Commun."; date: "2016-04-19"; volume: "7:11179. doi"; pages: " 
            10.1038/ncomms11179"
COMMENT     Sequence Label: pMLRT
FEATURES             Location/Qualifiers
     source          1..5745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..64
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        85..119
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             130..141
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             154..189
                     /codon_start=1
                     /product="6x(His-Asn) affinity tag"
                     /label=6xHN
                     /translation="HNHNHNHNHNHN"
     CDS             196..210
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     terminator      2190..2261
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2277..2304
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2357..2443
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2981..3931)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3979..4201
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4693..4787)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4818..5612)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4929..4948)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5539..5558
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tccgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
       61 aggccctttc gtcttcacct cgagttgaca gctagctcag tcctaggtac tgtgctagcg
      121 gaattcatta aagaggagaa aggtaccatg ggtcataatc ataatcataa tcataatcat
      181 aatcacaacg gtggagatga cgatgacaag ggtggtcgac aagcttggat ccctgcaggc
      241 ctcagggccc gatcgatggg accaatgggg cagcccctgc aagtgttgac cctaaatata
      301 gaagatgagt atcggctaca tgagacctca aaagagccag atgtttctct agggtccaca
      361 tggctgtctg attttcctca ggcctgggcg gaaaccgggg gcatgggact ggcagttcgc
      421 caagctcctc tgatcatacc tctgaaagca acctctaccc ccgtgtccat aaaacaatac
      481 cccatgtcac aagaagccag actggggatc aagccccaca tacagagact gttggaccag
      541 ggaatactgg taccctgcca gtccccctgg aacacgcccc tgctacccgt taagaaacca
      601 gggactaatg attataggcc tgtccaggat ctgagagaag tcaacaagcg ggtggaagac
      661 atccacccca ccgtgcccaa cccttacaac ctcttgagcg ggctcccacc gtcccaccag
      721 tggtacactg tgcttgattt aaaggatgcc tttttctgcc tgagactcca ccccaccagt
      781 cagcctctct tcgcctttga gtggagagat ccagagatgg gaatctcagg acaattgacc
      841 tggaccagac tcccacaggg tttcaaaaac agtcccaccc tgtttgatga ggcactgcac
      901 agagacctag cagacttccg gatccagcac ccagacttga tcctgctaca gtacgtggat
      961 gacttactgc tggccgccac ttctgagcta gactgccaac aaggtactcg ggccctgtta
     1021 caaaccctag ggaacctcgg gtatcgggcc tcggccaaga aagcccaaat ttgccagaaa
     1081 caggtcaagt atctggggta tcttctaaaa gagggtcaga gatggctgac tgaggccaga
     1141 aaagagactg tgatggggca gcctactccg aagacccctc gacaactaag ggagttccta
     1201 gggacggcag gcttctgtcg cctctggatc cctgggtttg cagaaatggc agcccccttg
     1261 taccctctca ccaaaacggg gactctgttt aattggggcc cagaccaaca aaaggcctat
     1321 caagaaatca agcaagctct tctaactgcc ccagccctgg ggttgccaga tttgactaag
     1381 ccctttgaac tctttgtcga cgagaagcag ggctacgcca aaggtgtcct aacgcaaaaa
     1441 ctgggacctt ggcgtcggcc ggtggcctac ctgtccaaaa agctagaccc agtagcagct
     1501 gggtggcccc cttgcctacg gatggtagca gccattgccg tactgacaaa ggatgcaggc
     1561 aagctaacca tgggacagcc actagtcatt ctggcccccc atgcagtaga ggcactagtc
     1621 aaacaacccc ccgaccgctg gctttccaac gcccggatga ctcactatca ggccttgctt
     1681 ttggacacgg accgggtcca gttcggaccg gtggtagccc tgaacccggc tacgctgctc
     1741 ccactgcctg aggaagggct gcaacacaac tgccttgata tcctggccga agcccacgga
     1801 acccgacccg acctaacgga ccagccgctc ccagacgccg accacacctg gtacacggat
     1861 ggaagcagtc tcttacaaga gggacagcgt aaggcgggag ctgcggtgac caccgagacc
     1921 gaggtaatct gggctaaagc cctgccagcc gggacatccg ctcagcgggc tgaactgata
     1981 gcactcaccc aggccctaaa gatggcagaa ggtaagaagc taaatgttta tactgatagc
     2041 cgttatgctt ttgctactgc ccatatccat ggagaaatat acagaaggcg tgggttgctc
     2101 acatcagaag gcaaagagat caaaaataaa gacgagatct ttaaatgacg ctgatagtgc
     2161 tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2221 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     2281 caccttcggg tgggcctttc tgcgtttata tataataaat gtccagacct gcaggcatgc
     2341 aagcctctag aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2401 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc cctagaccta
     2461 gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt gttatcagaa
     2521 tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcac tatttcttcc agaattgcca
     2581 tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat
     2641 aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct
     2701 caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg
     2761 tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttgaatgc
     2821 accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca
     2881 tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag
     2941 tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag
     3001 ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga
     3061 gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt
     3121 ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg
     3181 taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta
     3241 cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc
     3301 gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca
     3361 aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact
     3421 taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt
     3481 ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca
     3541 gattatattt tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa
     3601 ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc
     3661 ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag
     3721 ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag
     3781 tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca
     3841 cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt
     3901 catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat
     3961 cactatacca attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg
     4021 tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct
     4081 ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt
     4141 atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact
     4201 cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc
     4261 tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat
     4321 cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac
     4381 attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc
     4441 cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc
     4501 tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag
     4561 ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag
     4621 actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc
     4681 cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     4741 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc
     4801 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     4861 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     4921 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     4981 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     5041 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     5101 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     5161 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     5221 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     5281 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5341 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5401 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     5461 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5521 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5581 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     5641 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     5701 ttgcagggct tcccaacctt accagagggc gccccagctg gcaat
//
