LOCUS       sequence-382509-        8063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral golden gate cloning vector for expression of functional 
            Rhesus T-Cell Receptor genes with NGFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8063)
  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 8063)
  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: "10.2144/000114265";
            journalName: "Biotechniques"; date: "2015-03-1- 1"; volume: "58"; 
            issue: "3"; pages: "135-9"
COMMENT     Sequence Label: MSGV Rh Acceptor PGK-NGFR
FEATURES             Location/Qualifiers
     source          1..8063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     complement(2946..2969)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3335..3354
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     misc_feature    4283..4335
                     /label=MCS
                     /note="multiple cloning site"
     LTR             4378..4894
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5066..5082
                     /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(join(5090..5096,5097..5114,5115..5120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5135..5156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5273..5290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5444..6032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5524..5543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6203..6994,6995..7063))
                     /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     6826..6845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7064..7168)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7236..7254
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7292..7314)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7414..7433
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7627..7649
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7641..7658
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7642..7658
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7857..7876
                     /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 cctaaaatcc cgatcgttta 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 cccgaccccg ccgtgtatca gctgagaggc agcaagagca acgacaccag cgtgtgcctg
     1921 ttcaccgact tcgacagcgt gatgaacgtg tcccagagca aggacagcga cgtgcacatc
     1981 accgacaaga cagtgctgga catgcggagc atggacttca agagcaatgg cgccgtggcc
     2041 tggtccaaca agagcgactt tgcctgcacc agcgccttca aggactccgt gatccccgcc
     2101 gacacattct tcccaggcac cgagagcgtg tgcgacgcca acctggtgga aaagtccttc
     2161 gagacagaca tgaacctgaa cttccagaac ctgtccgtga tcggcttccg gatcctgctg
     2221 ctgaaagtgg ccggcttcaa cctgctgatg accctgagac tgtggtccag ccgggccaag
     2281 agatctggca gcggcgccac caatttcagc ctgctgaaac aggccgggga cgtggaagag
     2341 aaccctaagt cttctttttg aagacttgat ctgaagaagg tgttcccccc aaaggtggcc
     2401 gtgttcgagc catctgaggc cgagatctcc cacacccaga aagccaccct cgtgtgtctg
     2461 gccaccggct tctaccccga ccacgtggaa ctgtcttggt gggtcaacgg caaagaggtg
     2521 cacagcggcg tgtccaccga tccccagcct ctgaaagaac agcccgccct ggaagatagc
     2581 cggtactgcc tgagcagcag actgagagtg tccgccacct tctggcacaa cccccggaac
     2641 cacttcagat gccaggtgca gttctacggc ctgagcgagg acgacgagtg gaccgaggac
     2701 agagacaagc ctatcacaca gaaaatcagc gccgaagtgt ggggcagagc cgactgcggc
     2761 tttacaagcg tgtcctatca gcagggcgtg ctgagcgcca ccatcctgta cgagattctg
     2821 ctgggcaagg ccaccctgta cgccgtgctg gtgtctgccc tgatgctgat ggcaatggtc
     2881 aagcggaagg acttctagat ttaaattacc gggtagggga ggcgcttttc ccaaggcagt
     2941 ctggagcatg cgctttagca gccccgctgg cacttggcgc tacacaagtg gcctctggcc
     3001 tcgcacacat tccacatcca ccggtagcgc caaccggctc cgttctttgg tggccccttc
     3061 gcgccacttc tactcctccc ctagtcagga agtttccccc agcaagctcg cgtcgtgcag
     3121 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     3181 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgtt ccttcgcttt
     3241 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     3301 cgggcgggcg cccgaaggtc ctcccgaggc ccggcattct gcacgcttca aaagcgcacg
     3361 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacttgcag gcggccgcac
     3421 catgggggca ggtgccaccg gccgcgccat ggacgggccg cgcctgctgc tgttgctgct
     3481 tctgggggtg tcccttggag gtgccaagga ggcatgcccc acaggcctgt acacacacag
     3541 cggtgagtgc tgcaaagcct gcaacctggg cgagggtgtg gcccagcctt gtggagccaa
     3601 ccagaccgtg tgtgagccct gcctggacag cgtgacgttc tccgacgtgg tgagcgcgac
     3661 cgagccgtgc aagccgtgca ccgagtgcgt ggggctccag agcatgtcgg cgccatgcgt
     3721 ggaggccgac gacgccgtgt gccgctgcgc ctacggctac taccaggatg agacgactgg
     3781 gcgctgcgag gcgtgccgcg tgtgcgaggc gggctcgggc ctcgtgttct cctgccagga
     3841 caagcagaac accgtgtgcg aggagtgccc cgacggcacg tattccgacg aggccaacca
     3901 cgtggacccg tgcctgccct gcaccgtgtg cgaggacacc gagcgccagc tccgcgagtg
     3961 cacacgctgg gccgacgccg agtgcgagga gatccctggc cgttggatta cacggtccac
     4021 acccccagag ggctcggaca gcacagcccc cagcacccag gagcctgagg cacctccaga
     4081 acaagacctc atagccagca cggtggcagg tgtggtgacc acagtgatgg gcagctccca
     4141 gcccgtggtg acccgaggca ccaccgacaa cctcatccct gtctattgct ccatcctggc
     4201 tgctgtggtt gtgggtcttg tggcctacat agccttcaag aggtggaaca gctgagcggc
     4261 cgctacgtag atcgacggat ccctcgagct caagcttcga attctgcagt cgacggtacc
     4321 gcgggcccgg gatccgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     4381 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4441 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     4501 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4561 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     4621 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     4681 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     4741 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     4801 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     4861 gtgattgact acccgtcagc gggggtcttt cattgggtaa cagtttcttg aagttggaga
     4921 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
     4981 tgaatccaca tactccaata ctcctgaaat ccatcgatgg agttcattat ggacagcgca
     5041 gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag
     5101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5881 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7141 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     7201 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     7261 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     7321 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7381 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     7441 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     7501 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     7561 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     7621 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc cacgctctcc cttatgcgac
     7681 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     7741 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     7801 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     7861 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     7921 cgtccggcgt agaggcgatt taaagacagg atatcagtgg tccaggctct agttttgact
     7981 caacaatatc accagctgaa gcctatagag tacgagccat agataaaata aaagatttta
     8041 tttagtctcc agaaaaaggg ggg
//
