LOCUS       sequence-422378-        6629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral golden gate cloning vector for expression of functional 
            Human T-Cell Receptor genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6629)
  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 6629)
  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 Hu Acceptor
FEATURES             Location/Qualifiers
     source          1..6629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..515
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    559..611
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1764..1780)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     primer_bind     complement(2118..2134)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2150..2172)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     LTR             3012..3528
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3728..3747)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3875..3897)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4091..4110)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4210..4232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4270..4288)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4356..4460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4461..4529,4530..5321)
                     /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     complement(4679..4698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5492..6080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5981..6000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6234..6251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6368..6389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6404..6409,6410..6427,6428..6434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6442..6458
                     /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)."
ORIGIN
        1 atgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc aaggaacagc
       61 gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt acccgggcga
      121 cgcagtctat cggaggactg gcgcgccgag tgaggggttg tgggctcttt tattgagctc
      181 ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt tcaaataagg
      241 cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc tctagaaact
      301 gctgagggcg ggaccgcatc tggggaccat ctgttcttgg ccctgagccg gggcaggaac
      361 tgcttaccac agatatcctg tttggcccat attctgctgt ctctctgttc ctaaccttga
      421 tctgaacttc tctattctca gttatgtatt ttccatgcct tgcaaaatgg cgttacttaa
      481 gctagcttgc caaacctaca ggtggggtct ttcattcccc cctttttctg gagactaaat
      541 aaaatctttt attttatcgg atcccgggcc cgcggtaccg tcgactgcag aattcgaagc
      601 ttgagctcga gggatccgtc gactacaaat tcagaagtcc ttccgcttga ccatggccat
      661 cagcaccagg gcagacacca gcacggcgta cagggtggcc ttgcccagca ggatctcgta
      721 caggatggtg gcgctcagca cgccctgctg ataggacacg ctggtaaagc cgcaatcggc
      781 tctgccccag gcttcggcag acacgatctg ggtcacgggc ttggctctgt cctgggtcca
      841 ctcgtcgttc tcgctcaggc cgtagaactg cacctggcat ctgaagtggt tccgggggtt
      901 ctgccagaag gtggcggaca ctctcagtct gctgctcagg cagtaccggc tgtcgttcag
      961 ggcgggctgt tctttcagag gctggggatc ggtggacacg ccgctgtgca cctctttgcc
     1021 gttgacccac caagacagtt ccacgtggtc ggggaaaaag ccggtggcca ggcacacgag
     1081 ggtggctttc tgggtgtggc tgatctcggc ctcagaaggc tcgaacacgg ccacctctgg
     1141 ggggaacacc ttgttcagat caagtcttca aaaagaagac ttagggttct cttccacgtc
     1201 gccggcctgt ttcagcaggc tgaaattggt ggcgccgctg ccagatctct tggcccggct
     1261 ggaccacagt ctcagggtca tcagcaggtt gaagccggcc accttcagca gcaggatccg
     1321 gaagccgatc acgctcaggt tctggaagtt caggttggtg tctgtctcga agctcttttc
     1381 caccagcttc acgtcgcagc tgctctcggg gcttgggaag aatgtgtcct cggggataat
     1441 gctgttgttg aaggcgttgg cgcaggcgaa atcgctcttg ttggaccagg ccacggcgct
     1501 gttgctcttg aagtccatgc tccgcatgtc cagcacggtc ttgtcggtga tgtacacgtc
     1561 gctgtccttg ctctgggaca cgttggtctg gctgtcgaag tcggtgaaca ggcacacgct
     1621 cttgtcgctg ctcttgctgt cccgcagctg gtacacggca ggatcagggt tctgaagtct
     1681 tcaaaaaaag aagacttggt ggcttaatta acctgcaggg tctagaggat ggtccacccc
     1741 cggggtcggc agccttcacg tgggcggcgt gtatccaagc tgcgatgccg tctactttga
     1801 gggcggtggg ggtggtcagc aggactgtgt aaggtccttt ccagcgaggt tctaggttct
     1861 tagtctggtg tcggcggacc cacactgtgt cgccgactcg gtaagggtga ggtaccaccg
     1921 gtcggtccag ttgttcttgg taggctgccg ccagaggtct ccagacttcg tgctggacta
     1981 agtagagagc ctgtaagtga gcttggagag aggggctgtt agtaactctt gtcatgtcag
     2041 ggtcagggaa gtttacaagg ggcgggggtg ccccatataa gatctcatat ggccatatgg
     2101 gggcgcctag agaaggagtg agggctggat aaagggagga tcgaggcggg gtcgaacgag
     2161 gaggttcaag ggggagagac ggggcggatg gaggaagagg aggcggaggc ttagggtgta
     2221 caaagggctt gacccaggga ggggggtcaa aagccaaggc ttcccaggtc acgatgtagg
     2281 ggacctggtc tgggtgtcca tgcgggccag gtgaaaagac cttgatctta acctgggtga
     2341 tgaggtctcg gttaaaggtg ccgtctcgcg gccatccgac gttaaaggtt ggccattctg
     2401 cagagcagaa ggtaacccaa cgtctcttct tgacatctac cgactggttg tgagcgatcc
     2461 gctcgacatc tttccagtga cctaaggtca aacttaaggg agtggtaaca ggctagcccg
     2521 ggcccatatt ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc
     2581 tgcagcagac aagacgcgcg gcgcggcttc ggtcccaaac cgaaagcaaa aattcagacg
     2641 gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac cacatatccc tcctctaagg
     2701 ggggtgcacc aaagagtcct aaacgatcgg gattttagga ctcaggtcgg gccacaaaaa
     2761 cggcccccga agtccctggg acgtctccca gggttgcggc cgggtgttcc gaactcgtca
     2821 gttccaccac gggtccgcca gatacagagc tagttagcta actagtacag acgcaggcgc
     2881 aaacattaga tgccggcaca aacacgcaca aagacagaga cagacacgaa acgaccgctg
     2941 gccagcttac ctcccggcgg gggggtcggt ggtccctggg caggggtctc caaatctcgg
     3001 tggaacctcc aaatgaaaga cccccgaggt gggcagtcaa tcaatctgag gagaccctcc
     3061 caaggatcag cgagtccacg attcggatgc aaacagcaag aggctttatt gggaatacgg
     3121 gtacccgggc gacgcagtct atcggaggac tggcgcgccg agtgaggggt tgtgggctct
     3181 tttattgagc tcggggagca gaagcgcgcg aacagaagcg agaagcgaac tgattggtta
     3241 gttcaaataa ggcacagggt catttcaggt ccttggggca ccctggaaac atctgatggt
     3301 tctctagaaa ctgctgaggg cgggaccgca tctggggacc atctgttctt ggccctgagc
     3361 cggggcagga actgcttacc acagatatcc tgtttggccc atattctgct gtctctctgt
     3421 tcctaacctt gatctgaact tctctattct cagttatgta ttttccatgc cttgcaaaat
     3481 ggcgttactt aagctagctt gccaaaccta caggtggggt ctttcattcc cccctttttc
     3541 tggagactaa ataaaatctt ttattttatc tatggctcgt actctatagg cttcagctgg
     3601 tgatattgtt gagtcaaaac tagagcctgg accactgata tcctgtcttt aacaaattgg
     3661 actaatcgcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     3721 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     3781 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     3841 gccatctcct tgcatgcacc attccttgcg ccgtcgtttt acaacgtcgt gactgggaaa
     3901 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3961 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4021 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4081 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4141 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4201 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4261 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4321 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4681 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4981 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6241 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6301 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6361 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6421 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacaattct ccccagctct
     6481 ttctgcgctg tccataatga actccatcga tggatttcag gagtattgga gtatgtggat
     6541 tcaaaacagt ggctaattga gtcaggatta cttaatattc gactcctacc ctcagaatgt
     6601 tgttctccaa cttcaagaaa ctgttaccc
//
