LOCUS       Exported                8069 bp ds-DNA     circular SYN 23-JAN-2023
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 Jan 23, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8069
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1356..1378
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1394..1410
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1748..1764
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     promoter        2909..3407
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2944..2967)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3340..3359
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     misc_feature    4288..4340
                     /label=MCS
                     /note="multiple cloning site"
     LTR             4384..4898
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5070..5086
                     /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(5094..5124)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5139..5160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5277..5294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5448..6036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5528..5547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6207..7067)
                     /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     6830..6849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7068..7172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7240..7258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7296..7318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7418..7437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7631..7653
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7645..7662
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7646..7662
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7861..7880
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             8068..516
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttcgga
      721 acacccggcc gcaaccctgg gagacgtccc agggacttcg ggggccgttt ttgtggcccg
      781 acctgagtcc taaaatcccg atcgtttagg actctttggt gcacccccct tagaggaggg
      841 atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt
      901 gctttcggtt tgggaccgaa gccgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg
      961 ttgtctctgt ctgactgtgt ttctgtattt gtctgagaat atgggcccgg gctagcctgt
     1021 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     1081 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     1141 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     1201 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     1261 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     1321 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     1381 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata
     1441 tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg acatgacaag
     1501 agttactaac agcccctctc tccaagctca cttacaggct ctctacttag tccagcacga
     1561 agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg tggtacctca
     1621 cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga acctagaacc
     1681 tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca aagtagacgg
     1741 catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg gtggaccatc
     1801 ctctagaccg caacgttaat taagccacca agtcttcttt ttttgaagac ttcagaaccc
     1861 tgatcctgcc gtgtaccagc tgcgggacag caagagcagc gacaagagcg tgtgcctgtt
     1921 caccgacttc gacagccaga ccaacgtgtc ccagagcaag gacagcgacg tgtacatcac
     1981 cgacaagacc gtgctggaca tgcggagcat ggacttcaag agcaacagcg ccgtggcctg
     2041 gtccaacaag agcgatttcg cctgcgccaa cgccttcaac aacagcatta tccccgagga
     2101 cacattcttc ccaagccccg agagcagctg cgacgtgaag ctggtggaaa agagcttcga
     2161 gacagacacc aacctgaact tccagaacct gagcgtgatc ggcttccgga tcctgctgct
     2221 gaaggtggcc ggcttcaacc tgctgatgac cctgagactg tggtccagcc gggccaagag
     2281 atctggcagc ggcgccacca atttcagcct gctgaaacag gccggcgacg tggaagagaa
     2341 ccctaagtct tctttttgaa gacttgatct gaacaaggtg ttccccccag aggtggccgt
     2401 gttcgagcct tctgaggccg agatcagcca cacccagaaa gccaccctcg tgtgcctggc
     2461 caccggcttt ttccccgacc acgtggaact gtcttggtgg gtcaacggca aagaggtgca
     2521 cagcggcgtg tccaccgatc cccagcctct gaaagaacag cccgccctga acgacagccg
     2581 gtactgcctg agcagcagac tgagagtgtc cgccaccttc tggcagaacc cccggaacca
     2641 cttcagatgc caggtgcagt tctacggcct gagcgagaac gacgagtgga cccaggacag
     2701 agccaagccc gtgacccaga tcgtgtctgc cgaagcctgg ggcagagccg attgcggctt
     2761 taccagcgtg tcctatcagc agggcgtgct gagcgccacc atcctgtacg agatcctgct
     2821 gggcaaggcc accctgtacg ccgtgctggt gtctgccctg gtgctgatgg ccatggtcaa
     2881 gcggaaggac ttctgaattt aaattaccgg gtaggggagg cgcttttccc aaggcagtct
     2941 ggagcatgcg ctttagcagc cccgctggca cttggcgcta cacaagtggc ctctggcctc
     3001 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     3061 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     3121 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     3181 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     3241 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     3301 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     3361 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcaggcggc
     3421 cgcaccatgg gggcaggtgc cactggccgc gccatggacg ggccgcgcct gctgctgttg
     3481 ctgcttctgg gggtgtccct tggaggtgcc aaggaggcat gccccacagg cctgtacaca
     3541 cacagcggtg agtgctgcaa agcctgcaac ctgggcgagg gtgtggccca gccttgtgga
     3601 gccaaccaga ccgtgtgtga gccctgcctg gacagcgtga cgttctccga cgtggtgagc
     3661 gcgaccgagc cgtgcaagcc gtgcaccgag tgcgtggggc tccagagcat gtcggcgcca
     3721 tgcgtggagg ccgacgacgc cgtgtgccgc tgcgcctacg gctactacca ggatgagacg
     3781 actgggcgct gcgaggcgtg ccgcgtgtgc gaggcgggct cgggcctcgt gttctcctgc
     3841 caggacaagc agaacaccgt gtgcgaggag tgccccgacg gcacgtattc cgacgaggcc
     3901 aaccacgtgg acccgtgcct gccctgcacc gtgtgcgagg acaccgagcg ccagctccgc
     3961 gagtgcacac gctgggccga cgccgagtgc gaggagatcc ctggccgttg gattacacgg
     4021 tccacacccc cagagggctc ggacagcaca gcccccagca cccaggagcc tgaggcacct
     4081 ccagaacaag acctcatagc cagcacggtg gcaggtgtgg tgaccacagt gatgggcagc
     4141 tcccagcccg tggtgacccg aggcaccacc gacaacctca tccctgtcta ttgctccatc
     4201 ctggctgctg tggttgtggg tcttgtggcc tacatagcct tcaagaggtg gaacagctga
     4261 gcggccgcta cgtagatcga cggatccctc gagctcaagc ttcgaattct gcagtcgacg
     4321 gtaccgcggg cccgggatcc gataaaataa aagattttat ttagtctcca gaaaaagggg
     4381 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4441 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     4501 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4561 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     4621 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4681 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4741 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     4801 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     4861 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     4921 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     4981 gttttgaatc cacatactcc aatactcctg aaatccatcg atggagttca ttatggacag
     5041 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     5101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5881 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7201 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7261 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7321 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7381 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     7441 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     7501 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7561 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7621 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccacgct ctcccttatg
     7681 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     7741 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     7801 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     7861 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     7921 gatgcgtccg gcgtagaggc gatttaaaga caggatatca gtggtccagg ctctagtttt
     7981 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     8041 tttatttagt ctccagaaaa aggggggaa
//
