LOCUS       Exported                6699 bp ds-DNA     circular SYN 23-JAN-2023
DEFINITION  Retroviral golden gate cloning vector for expression of functional 
            Human T-Cell Receptor genes.
ACCESSION   .
VERSION     .
KEYWORDS    MSGV Hu Acceptor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6699)
  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 6699)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 23, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6699
                     /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"
     misc_feature    2920..2972
                     /label=MCS
                     /note="multiple cloning site"
     LTR             3016..3530
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3702..3718
                     /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(3726..3756)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3771..3792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3909..3926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4080..4668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4160..4179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4839..5699)
                     /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     5462..5481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5700..5804)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5872..5890
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5928..5950)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6050..6069
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6263..6285
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6277..6294
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6278..6294
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6493..6512
                     /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 cctgcaggtt aattaagcca ccaagtcttc tttttttgaa gacttcagaa
     1861 ccctgatcct gccgtgtacc agctgcggga cagcaagagc agcgacaaga gcgtgtgcct
     1921 gttcaccgac ttcgacagcc agaccaacgt gtcccagagc aaggacagcg acgtgtacat
     1981 caccgacaag accgtgctgg acatgcggag catggacttc aagagcaaca gcgccgtggc
     2041 ctggtccaac aagagcgatt tcgcctgcgc caacgccttc aacaacagca ttatccccga
     2101 ggacacattc ttcccaagcc ccgagagcag ctgcgacgtg aagctggtgg aaaagagctt
     2161 cgagacagac accaacctga acttccagaa cctgagcgtg atcggcttcc ggatcctgct
     2221 gctgaaggtg gccggcttca acctgctgat gaccctgaga ctgtggtcca gccgggccaa
     2281 gagatctggc agcggcgcca ccaatttcag cctgctgaaa caggccggcg acgtggaaga
     2341 gaaccctaag tcttcttttt gaagacttga tctgaacaag gtgttccccc cagaggtggc
     2401 cgtgttcgag ccttctgagg ccgagatcag ccacacccag aaagccaccc tcgtgtgcct
     2461 ggccaccggc tttttccccg accacgtgga actgtcttgg tgggtcaacg gcaaagaggt
     2521 gcacagcggc gtgtccaccg atccccagcc tctgaaagaa cagcccgccc tgaacgacag
     2581 ccggtactgc ctgagcagca gactgagagt gtccgccacc ttctggcaga acccccggaa
     2641 ccacttcaga tgccaggtgc agttctacgg cctgagcgag aacgacgagt ggacccagga
     2701 cagagccaag cccgtgaccc agatcgtgtc tgccgaagcc tggggcagag ccgattgcgg
     2761 ctttaccagc gtgtcctatc agcagggcgt gctgagcgcc accatcctgt acgagatcct
     2821 gctgggcaag gccaccctgt acgccgtgct ggtgtctgcc ctggtgctga tggccatggt
     2881 caagcggaag gacttctgaa tttgtagatc gacggatccc tcgagctcaa gcttcgaatt
     2941 ctgcagtcga cggtaccgcg ggcccgggat ccgataaaat aaaagatttt atttagtctc
     3001 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     3061 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     3121 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3181 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     3241 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3301 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3361 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     3421 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     3481 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt
     3541 ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact
     3601 caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatccat cgatggagtt
     3661 cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt
     3721 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3781 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3841 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3901 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3961 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4021 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4081 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4141 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4201 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4261 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4321 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4381 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4441 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4501 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4561 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4621 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4681 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4741 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4801 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4861 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4921 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4981 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5041 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5101 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5161 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5221 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5281 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5341 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5401 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5461 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5521 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5581 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5641 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5701 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5761 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5821 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5881 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5941 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     6001 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     6061 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     6121 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6181 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     6241 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccacg
     6301 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     6361 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     6421 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     6481 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     6541 gatgccggcc acgatgcgtc cggcgtagag gcgatttaaa gacaggatat cagtggtcca
     6601 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
     6661 aaaataaaag attttattta gtctccagaa aaagggggg
//
