LOCUS       Exported                4975 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  gene cloning of MR1A.
ACCESSION   .
VERSION     .
KEYWORDS    pHMR1A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4975)
  AUTHORS   Lepore M, Kalinichenko A, Colone A, Paleja B, Singhal A, Tschumi A, 
            Lee B, Poidinger M, Zolezzi F, Quagliata L, Sander P, Newell E, 
            Bertoletti A, Terracciano L, De Libero G, Mori L
  TITLE     Parallel T-cell cloning and deep sequencing of human MAIT cells 
            reveal stable oligoclonal TCRbeta repertoire.
  JOURNAL   Nat Commun. 2014 May 15;5:3866. doi: 10.1038/ncomms4866.
  PUBMED    24832684
REFERENCE   2  (bases 1 to 4975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4975
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1207..1223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1207..1223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1220..1242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1231..1247
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1255..1285)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1300..1321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1438..1455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1609..2197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1689..1708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2368..3228)
                     /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     2991..3010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3246..4040)
                     /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(3357..3376)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3967..3986
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(4384..4812)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4494..4515)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4706..4725
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4938..4960
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4952..4969
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4953..4969
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc cctctagatg catgctcgag cggccgccag
       61 tgtgatggat atctgcagaa ttcgccctta tctcgaggga ctatggggga actgatggcg
      121 ttcctgttac ctctcatcat tgtgttaatg gtgaagcaca gcgattcccg gacgcactct
      181 ctgagatatt ttcgcctggg cgtttcggat cccatccatg gggtccctga atttatttcg
      241 gttgggtacg tggactcgca ccctatcacc acatatgaca gtgtcactcg gcagaaggag
      301 ccacgggccc catggatggc agagaacctc gcgcctgatc actgggagag gtacactcag
      361 ctgctgaggg gctggcagca gatgttcaag gtggaactga agcgcctaca gaggcactac
      421 aatcactcag ggtctcacac ttaccagaga atgattggct gtgagctgct ggaggatgga
      481 agcaccacag gatttctgca gtatgcatat gacgggcagg atttcctgat cttcaataaa
      541 gacaccctct cctggctggc tgtagataat gtggctcaca ccatcaagca ggcatgggag
      601 gccaatcagc atgagttgct gtatcaaaag aattggctgg aagaagaatg tattgcctgg
      661 ctaaagagat tcctggagta tgggaaagac accctacaaa gaacagagcc cccactggtc
      721 agagtaaatc gcaaagaaac ttttccaggg gttacagctc tcttctgcaa agctcatggc
      781 ttttaccccc cagaaattta catgacatgg atgaaaaacg gggaagaaat tgtccaagaa
      841 attgattatg gagacattct tcccagtggg gatggaacct atcaggcgtg ggcatcaatt
      901 gagcttgatc ctcagagcag caacctttac tcctgtcatg tggagcactg cggtgtccac
      961 atggttcttc aggtccccca ggaatcagaa actatccctc ttgtgatgaa agctgtctct
     1021 gggtccattg tccttgtcat tgtgctggct ggagttggtg ttctagtctg gagaagaagg
     1081 ccccgagagc aaaatggagc catctacctt ccaacaccag atcgaaagct tttaagggcg
     1141 aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttggcgta
     1201 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1261 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1321 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3241 tcaattcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     3301 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     3361 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     3421 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     3481 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     3541 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     3601 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     3661 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     3721 aggtgggatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     3781 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     3841 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     3901 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     3961 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     4021 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat
     4081 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
     4141 ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc
     4201 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
     4261 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     4321 tgcggactgg ctttctacgt gttccgcttc ctttagcagc ccttgcgccc tgaattttgt
     4381 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4441 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4501 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4561 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4621 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4681 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4741 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4801 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4861 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4921 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattg
//
