LOCUS Exported 6704 bp ds-DNA circular SYN 23-JAN-2023 DEFINITION Retroviral golden gate cloning vector for expression of functional Rhesus Macaque T-Cell Receptor genes. ACCESSION . VERSION . KEYWORDS MSGV Rh Acceptor 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 Jan 23, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6704 /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" 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(3729..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(4842..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 6703..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 //