LOCUS       sequence-382507-        6704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral golden gate cloning vector for expression of functional 
            Rhesus Macaque T-Cell Receptor genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  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 6704)
  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
FEATURES             Location/Qualifiers
     source          1..6704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     misc_feature    2923..2975
                     /label=MCS
                     /note="multiple cloning site"
     LTR             3019..3533
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3705..3721
                     /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(3729..3735,3736..3753,3754..3759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3774..3795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3912..3929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4083..4671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4163..4182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4842..5633,5634..5702))
                     /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     5465..5484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5703..5807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5875..5893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5931..5953)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6053..6072
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6266..6288
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6280..6297
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6281..6297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6496..6515
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6703..6704,1..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 ccatgttaat taagccacca agtcttcttt ttttgaagac ttcagaaccc
     1861 cgaccccgcc gtgtatcagc tgagaggcag caagagcaac gacaccagcg tgtgcctgtt
     1921 caccgacttc gacagcgtga tgaacgtgtc ccagagcaag gacagcgacg tgcacatcac
     1981 cgacaagaca gtgctggaca tgcggagcat ggacttcaag agcaatggcg ccgtggcctg
     2041 gtccaacaag agcgactttg cctgcaccag cgccttcaag gactccgtga tccccgccga
     2101 cacattcttc ccaggcaccg agagcgtgtg cgacgccaac ctggtggaaa agtccttcga
     2161 gacagacatg aacctgaact tccagaacct gtccgtgatc ggcttccgga tcctgctgct
     2221 gaaagtggcc ggcttcaacc tgctgatgac cctgagactg tggtccagcc gggccaagag
     2281 atctggcagc ggcgccacca atttcagcct gctgaaacag gccggggacg tggaagagaa
     2341 ccctaagtct tctttttgaa gacttgatct gaagaaggtg ttccccccaa aggtggccgt
     2401 gttcgagcca tctgaggccg agatctccca cacccagaaa gccaccctcg tgtgtctggc
     2461 caccggcttc taccccgacc acgtggaact gtcttggtgg gtcaacggca aagaggtgca
     2521 cagcggcgtg tccaccgatc cccagcctct gaaagaacag cccgccctgg aagatagccg
     2581 gtactgcctg agcagcagac tgagagtgtc cgccaccttc tggcacaacc cccggaacca
     2641 cttcagatgc caggtgcagt tctacggcct gagcgaggac gacgagtgga ccgaggacag
     2701 agacaagcct atcacacaga aaatcagcgc cgaagtgtgg ggcagagccg actgcggctt
     2761 tacaagcgtg tcctatcagc agggcgtgct gagcgccacc atcctgtacg agattctgct
     2821 gggcaaggcc accctgtacg ccgtgctggt gtctgccctg atgctgatgg caatggtcaa
     2881 gcggaaggac ttctagcggc cgctacgtag atcgacggat ccctcgagct caagcttcga
     2941 attctgcagt cgacggtacc gcgggcccgg gatccgataa aataaaagat tttatttagt
     3001 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     3061 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
     3121 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3181 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
     3241 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3301 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     3361 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
     3421 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     3481 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac
     3541 agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg
     3601 actcaattag ccactgtttt gaatccacat actccaatac tcctgaaatc catcgatgga
     3661 gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta tccgctcaca
     3721 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3781 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3841 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3901 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3961 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4021 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4141 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4261 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4501 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4561 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4621 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4681 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4741 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4801 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4861 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4921 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4981 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5041 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5101 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5161 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5221 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5281 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5341 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5401 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5461 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5521 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5581 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5641 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5701 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5761 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5821 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5881 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5941 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6001 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     6061 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     6121 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     6301 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6361 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6421 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6481 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6541 ggtgatgccg gccacgatgc gtccggcgta gaggcgattt aaagacagga tatcagtggt
     6601 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     6661 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaa
//
