LOCUS       MSGV Hu Acceptor PGK-NGFR             8069 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Retroviral golden gate cloning vector for expression of functional 
            Human T-Cell Receptor genes with NGFR.
ACCESSION   .
VERSION     .
KEYWORDS    MSGV Hu Acceptor PGK-NGFR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  AUTHORS   Coren LV, Jain S, Trivett MT, Ohlen C, Ott DE
  TITLE     Production of retroviral constructs for effective transfer and 
            expression of T-cell receptor genes using Golden Gate cloning.
  JOURNAL   Biotechniques. 2015 Mar 1;58(3):135-9. doi: 10.2144/000114265. 
            eCollection 2015.
  PUBMED    25757546
REFERENCE   2  (bases 1 to 8069)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8069
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..518
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     1358..1380
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1396..1412
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1750..1766
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     promoter        2911..3409
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2946..2969)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3342..3361
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     misc_feature    4290..4342
                     /label=MCS
                     /note="multiple cloning site"
     LTR             4386..4900
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5072..5088
                     /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(5096..5126)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5141..5162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5279..5296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5450..6038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5530..5549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6209..7069)
                     /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     6832..6851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7070..7174)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7242..7260
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7298..7320)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7420..7439
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7633..7655
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7647..7664
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7648..7664
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7863..7882
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaaatga ccctgtgcct tatttgaact aaccaatcag
      301 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
      361 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
      421 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
      481 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
      541 agacccctgc ctagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
      601 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
      661 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg
      721 gaacacccgg ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc
      781 cgacctgagt ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag
      841 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
      901 ttgctttcgg tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg
      961 tgttgtctct gtctgactgt gtttctgtat ttgtctgaga atatgggccc gggctagcct
     1021 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
     1081 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
     1141 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
     1201 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
     1261 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
     1321 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
     1381 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgcccc catatggcca
     1441 tatgagatct tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca
     1501 agagttacta acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac
     1561 gaagtctgga gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct
     1621 cacccttacc gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa
     1681 cctcgctgga aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac
     1741 ggcatcgcag cttggataca cgccgcccac gtgaaggctg ccgaccccgg gggtggacca
     1801 tcctctagac cgcaacgtta attaagccac caagtcttct ttttttgaag acttcagaac
     1861 cctgatcctg ccgtgtacca gctgcgggac agcaagagca gcgacaagag cgtgtgcctg
     1921 ttcaccgact tcgacagcca gaccaacgtg tcccagagca aggacagcga cgtgtacatc
     1981 accgacaaga ccgtgctgga catgcggagc atggacttca agagcaacag cgccgtggcc
     2041 tggtccaaca agagcgattt cgcctgcgcc aacgccttca acaacagcat tatccccgag
     2101 gacacattct tcccaagccc cgagagcagc tgcgacgtga agctggtgga aaagagcttc
     2161 gagacagaca ccaacctgaa cttccagaac ctgagcgtga tcggcttccg gatcctgctg
     2221 ctgaaggtgg ccggcttcaa cctgctgatg accctgagac tgtggtccag ccgggccaag
     2281 agatctggca gcggcgccac caatttcagc ctgctgaaac aggccggcga cgtggaagag
     2341 aaccctaagt cttctttttg aagacttgat ctgaacaagg tgttcccccc agaggtggcc
     2401 gtgttcgagc cttctgaggc cgagatcagc cacacccaga aagccaccct cgtgtgcctg
     2461 gccaccggct ttttccccga ccacgtggaa ctgtcttggt gggtcaacgg caaagaggtg
     2521 cacagcggcg tgtccaccga tccccagcct ctgaaagaac agcccgccct gaacgacagc
     2581 cggtactgcc tgagcagcag actgagagtg tccgccacct tctggcagaa cccccggaac
     2641 cacttcagat gccaggtgca gttctacggc ctgagcgaga acgacgagtg gacccaggac
     2701 agagccaagc ccgtgaccca gatcgtgtct gccgaagcct ggggcagagc cgattgcggc
     2761 tttaccagcg tgtcctatca gcagggcgtg ctgagcgcca ccatcctgta cgagatcctg
     2821 ctgggcaagg ccaccctgta cgccgtgctg gtgtctgccc tggtgctgat ggccatggtc
     2881 aagcggaagg acttctgaat ttaaattacc gggtagggga ggcgcttttc ccaaggcagt
     2941 ctggagcatg cgctttagca gccccgctgg cacttggcgc tacacaagtg gcctctggcc
     3001 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     3061 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     3121 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     3181 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     3241 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     3301 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     3361 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcaggcg
     3421 gccgcaccat gggggcaggt gccactggcc gcgccatgga cgggccgcgc ctgctgctgt
     3481 tgctgcttct gggggtgtcc cttggaggtg ccaaggaggc atgccccaca ggcctgtaca
     3541 cacacagcgg tgagtgctgc aaagcctgca acctgggcga gggtgtggcc cagccttgtg
     3601 gagccaacca gaccgtgtgt gagccctgcc tggacagcgt gacgttctcc gacgtggtga
     3661 gcgcgaccga gccgtgcaag ccgtgcaccg agtgcgtggg gctccagagc atgtcggcgc
     3721 catgcgtgga ggccgacgac gccgtgtgcc gctgcgccta cggctactac caggatgaga
     3781 cgactgggcg ctgcgaggcg tgccgcgtgt gcgaggcggg ctcgggcctc gtgttctcct
     3841 gccaggacaa gcagaacacc gtgtgcgagg agtgccccga cggcacgtat tccgacgagg
     3901 ccaaccacgt ggacccgtgc ctgccctgca ccgtgtgcga ggacaccgag cgccagctcc
     3961 gcgagtgcac acgctgggcc gacgccgagt gcgaggagat ccctggccgt tggattacac
     4021 ggtccacacc cccagagggc tcggacagca cagcccccag cacccaggag cctgaggcac
     4081 ctccagaaca agacctcata gccagcacgg tggcaggtgt ggtgaccaca gtgatgggca
     4141 gctcccagcc cgtggtgacc cgaggcacca ccgacaacct catccctgtc tattgctcca
     4201 tcctggctgc tgtggttgtg ggtcttgtgg cctacatagc cttcaagagg tggaacagct
     4261 gagcggccgc tacgtagatc gacggatccc tcgagctcaa gcttcgaatt ctgcagtcga
     4321 cggtaccgcg ggcccgggat ccgataaaat aaaagatttt atttagtctc cagaaaaagg
     4381 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     4441 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     4501 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     4561 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     4621 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     4681 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     4741 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     4801 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     4861 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
     4921 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
     4981 ctgttttgaa tccacatact ccaatactcc tgaaatccat cgatggagtt cattatggac
     5041 agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca
     5101 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5161 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5221 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5881 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7081 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7141 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7201 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7261 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7321 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7381 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7441 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     7501 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     7561 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     7621 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccacg ctctccctta
     7681 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     7741 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     7801 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     7861 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     7921 acgatgcgtc cggcgtagag gcgatttaaa gacaggatat cagtggtcca ggctctagtt
     7981 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     8041 attttattta gtctccagaa aaagggggg
//