LOCUS       sequence-222068-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  translational initiation factor 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  AUTHORS   Villarreal F, Contreras-Llano LE, Chavez M, Ding Y, Fan J, Pan T, 
            Tan C
  TITLE     Synthetic microbial consortia enable rapid assembly of pure 
            translation machinery.
  JOURNAL   Nat Chem Biol. 2018 Jan;14(1):29-35. doi: 10.1038/nchembio.2514. 
            Epub 2017 Nov 13.
  PUBMED    29131146
REFERENCE   2  (bases 1 to 6386)
  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"; doi: 
            "10.1038/nchembio.2514"; journalName: "Nat Chem Biol"; date: 
            "2018-01"; volume: "14"; issue: "1"; pages: "29-35"
COMMENT     Sequence Label: B3-1
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..91
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        72..90
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             130..152
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             162..179
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2926..2944)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2940..2987
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3596..4546)
                     /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      4594..4816
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5308..5402)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5433..6227)
                     /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(5544..5563)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6154..6173
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 catgacatta acctataaaa ataggcggat cacgaggccc tttcgtcttc acctcgatga
       61 tcccgcgaaa ttaatacgac tcactatagg ggaattgaga gcggataaca attcccctct
      121 agaaataatt ttgtttaact ttaagaagga gatataccat gcatcatcat caccatcatt
      181 taacagatgt aacgattaaa acgctggccg cagagcgaca gacctccgtg gaacgcctgg
      241 tacagcaatt tgctgatgca ggtatccgga agtctgctga cgactctgtg tctgcacaag
      301 agaaacagac tttgattgac cacctgaatc agaaaaattc aggcccggac aaattgacgc
      361 tgcaacgtaa aacacgcagc acccttaaca ttcctggtac cggtggaaaa agcaaatcgg
      421 tacaaatcga agtccgcaag aaacgcacct ttgtgaaacg cgatccgcaa gaggctgaac
      481 gccttgcagc ggaagagcaa gcgcagcgtg aagcggaaga gcaagcccgt cgtgaggcag
      541 aagaatcggc taaacgcgag gcgcaacaaa aagctgaacg tgaggccgca gaacaagcta
      601 agcgtgaagc tgctgaacaa gcgaaacgtg aagctgcgga aaaagacaaa gtgagcaatc
      661 aacaagacga tatgactaaa aacgcccagg ctgaaaaagc ccgccgtgag caggaagctg
      721 cagagctcaa gcgtaaagct gaagaagaag cgcgtcgtaa actcgaagaa gaagcacgtc
      781 gcgttgctga agaagcacgt cgtatggcgg aagaaaacaa atggactgat aacgcggaac
      841 cgactgaaga ttccagcgat tatcacgtca ctacttctca acatgctcgc caggcagaag
      901 acgaaagcga tcgtgaagtc gaaggcggcc gtggccgtgg tcgtaacgcg aaagcagcgc
      961 gtccgaagaa aggcaacaaa cacgctgaat caaaagctga tcgtgaagaa gcacgcgcag
     1021 cagtacgtgg cggtaaaggc ggaaaacgta aaggttcttc gctgcagcaa ggcttccaga
     1081 agcctgctca ggccgttaac cgtgacgttg tgatcggcga aactatcacc gttggcgaac
     1141 tggcgaacaa gatggcggtt aaaggctctc aggtcatcaa agcgatgatg aaactgggcg
     1201 caatggcaac catcaaccag gttatcgatc aggaaaccgc acagctggtt gctgaagaga
     1261 tgggccataa agttatcctg cgtcgtgaaa acgagctgga agaggcggta atgagcgacc
     1321 gtgacacggg tgctgcggct gaaccgcgcg cgccggttgt gaccatcatg ggtcacgttg
     1381 accacggtaa aacctctctg ctggactaca ttcgttcaac gaaagtggcc tctggcgaag
     1441 cgggcggcat tacccagcac attggtgcat accacgttga aactgaaaac ggcatgatca
     1501 ccttcctgga caccccgggg cacgccgcgt ttacttcaat gcgtgctcgt ggtgcgcagg
     1561 caacggacat cgtagtcctg gttgttgctg ccgacgacgg tgtgatgccg cagaccatcg
     1621 aagcaatcca gcacgcgaaa gcggcgcagg taccggtggt ggttgcagtg aacaagatcg
     1681 ataaaccaga agctgatccg gatcgcgtta agaacgaact ctcccagtac ggcatcctgc
     1741 cggaagagtg gggcggtgaa agccagttcg tacacgtatc tgcgaaagcg ggtaccggta
     1801 tcgatgaact gctggacgct atcctgctgc aggcggaagt tctggagctg aaagcggtac
     1861 gtaaaggtat ggcgagcggt gcggttatcg aatccttcct cgataaaggt cgtggtccgg
     1921 ttgctaccgt tctggtacgt gaaggtactc tgcacaaggg cgatatcgtt ctgtgtggct
     1981 tcgaatacgg tcgtgttcgt gcgatgcgta acgaactggg tcaggaagtg ctggaagcgg
     2041 gtccgtccat tccggtggaa atcctcggcc tgtccggcgt accggctgcg ggtgatgaag
     2101 ttaccgttgt acgtgacgag aagaaagcgc gtgaagttgc actctatcgt cagggtaaat
     2161 tccgcgaagt taaactggcg cgtcagcaga aatctaaact cgagaacatg ttcgccaaca
     2221 tgaccgaagg cgaagttcac gaagtgaata tcgtcctgaa ggcagacgta cagggttctg
     2281 tcgaagcgat ctccgactcc ttgctgaaac tgtctactga cgaagttaaa gtgaagatca
     2341 tcggttctgg cgtaggtggt atcaccgaaa ccgacgccac cctggctgcg gcgtccaacg
     2401 ccatcctggt tggctttaac gtacgtgctg atgcctctgc acgtaaagtg attgaagcgg
     2461 aaagcctgga tctgcgttac tactccgtca tctataacct gattgacgaa gtgaaagcgg
     2521 cgatgagcgg tatgctgtct ccggaactga aacagcagat tatcggtctg gcggaagttc
     2581 gtgacgtgtt caaatcgccg aaatttggtg ccatcgcagg ctgtatggtt accgaaggtg
     2641 tggttaaacg tcacaacccg atccgcgttc tgcgtgacaa cgtggttatc tacgaaggcg
     2701 agctggagtc cctgcgccgc ttcaaagatg acgttaacga agtccgtaac ggtatggaat
     2761 gtggtatcgg cgttaagaac tacaacgacg tccgcactgg cgatgtgatc gaagtattcg
     2821 aaatcatcga gatccaacgt accattgctt attaaagcag aggccatatg ctcgaggatc
     2881 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     2941 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     3001 ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac
     3061 agcatccagg gcctagggta cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct
     3121 agtttgttat cagaatcgca gatccggctt cagccggttt gccggctgaa agcgctattt
     3181 cttccagaat tgccatgatt ttttccccac gggaggcgtc actggctccc gtgttgtcgg
     3241 cagctttgat tcgataagca gcatcgcctg tttcaggctg tctatgtgtg actgttgagc
     3301 tgtaacaagt tgtctcaggt gttcaatttc atgttctagt tgctttgttt tactggtttc
     3361 acctgttcta ttaggtgtta catgctgttc atctgttaca ttgtcgatct gttcatggtg
     3421 aacagctttg aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc
     3481 gttttcatct gtgcatatgg acagttttcc ctttgatatg taacggtgaa cagttgttct
     3541 acttttgttt gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga
     3601 tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg
     3661 agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt gcctcaaaac
     3721 tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta
     3781 ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt
     3841 tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat
     3901 cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt
     3961 tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa
     4021 gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct
     4081 tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag
     4141 acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata
     4201 tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact
     4261 ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct
     4321 ctctggttgc tttagctaat acaccataag cattttccct actgatgttc atcatctgag
     4381 cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg
     4441 ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc
     4501 gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt
     4561 ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc
     4621 ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa
     4681 attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc
     4741 aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca
     4801 gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa
     4861 cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg
     4921 caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc
     4981 tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa
     5041 tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag
     5101 cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt
     5161 tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg
     5221 tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt
     5281 acccgtctta ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     5341 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     5401 tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     5461 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     5521 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     5581 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     5641 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     5701 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     5761 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     5821 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     5881 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     5941 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     6001 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     6061 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     6121 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     6181 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     6241 catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     6301 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     6361 tccgacgtct aagaaaccat tattat
//
