LOCUS       Exported                7856 bp ds-DNA     circular SYN 10-NOV-2022
DEFINITION  a bicistronic IRES-GFP containing retroviral vector, MCS is expanded
            from pMSCV-IRES-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7856)
  AUTHORS   Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ, 
            Vignali DA
  TITLE     Generation of T-cell receptor retrogenic mice.
  JOURNAL   Nat Protoc. 2006;1(1):406-17.
  PUBMED    17406263
REFERENCE   2  (bases 1 to 7856)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 14, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7856
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     gene            1413..2786
                     /label=Cd4
     misc_feature    2793..3369
                     /label=IRES
     primer_bind     complement(2962..2979)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3189..3208
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3370..4089
                     /label=mAmetrine 1.1
                     /label=mAmetrine\1.1
                     /note="/vntifkey=21"
     primer_bind     complement(3415..3436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3676..3695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4023..4043
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4251..4765
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4927..4943
                     /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(4951..4981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4996..5017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5134..5151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5305..5893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5385..5404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6064..6924)
                     /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     6687..6706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6925..7029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7097..7115
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7153..7175)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7275..7294
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7488..7510
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7638..7657
                     /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 ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tcatgtgccg agccatctct cttaggcgct
     1441 tgctgctgct gctgctgcag ctgtcacaac tcctagctgt cactcaaggg aagacgctgg
     1501 tgctggggaa ggaaggggaa tcagcagaac tgccctgcga gagttcccag aagaagatca
     1561 cagtcttcac ctggaagttc tctgaccaga ggaagattct ggggcagcat ggcaaaggtg
     1621 tattaattag aggaggttcg ccttcgcagt ttgatcgttt tgattccaaa aaaggggcat
     1681 gggagaaagg atcgtttcct ctcatcatca ataaacttaa gatggaagac tctcagactt
     1741 atatctgtga gctggagaac aggaaagagg aggtggagtt gtgggtgttc aaagtgacct
     1801 tcagtccggg taccagcctg ttgcaagggc agagcctgac cctgaccttg gatagcaact
     1861 ctaaggtctc taaccccttg acagagtgca aacacaaaaa gggtaaagtt gtcagtggtt
     1921 ccaaagttct ctccatgtcc aacctaaggg ttcaggacag cgacttctgg aactgcaccg
     1981 tgaccctgga ccagaaaaag aactggttcg gcatgacact ctcagtgctg ggttttcaga
     2041 gcacagctat cacggcctat aagagtgagg gagagtcagc ggagttctcc ttcccactca
     2101 actttgcaga ggaaaacggg tggggagagc tgatgtggaa ggcagagaag gattctttct
     2161 tccagccctg gatctccttc tccataaaga acaaagaggt gtccgtacaa aagtccacca
     2221 aagacctcaa gctccagctg aaggaaacgc tcccactcac cctcaagata ccccaggtct
     2281 cgcttcagtt tgctggttct ggcaacctga ctctgactct ggacaaaggg acactgcatc
     2341 aggaagtgaa cctggtggtg atgaaagtgg ctcagctcaa caatactttg acctgtgagg
     2401 tgatgggacc tacctctccc aagatgagac tgaccctgaa gcaggagaac caggaggcca
     2461 gggtctctga ggagcagaaa gtagttcaag tggtggcccc tgagacaggg ctgtggcagt
     2521 gtctactgag tgaaggtgat aaggtcaaga tggactccag gatccaggtt ttatccagag
     2581 gggtgaacca gacagtgttc ctggcttgcg tgctgggtgg ctccttcggc tttctgggtt
     2641 tccttgggct ctgcatcctc tgctgtgtca ggtgccggca ccaacagcgc caggcagcac
     2701 gaatgtctca gatcaagagg ctcctcagtg agaagaagac ctgccagtgc ccccaccgga
     2761 tgcagaagag ccataatctc atctgactcg agcgggatca attccgcccc ccccctaacg
     2821 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     2881 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2941 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3001 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     3061 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3121 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3181 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3241 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3301 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3361 gataatacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3421 gagctggacg gcgacgtaaa cggccacagg ttcagcgtgc gcggcgaggg cgagggcgat
     3481 gccaccaacg gcaagctgac cctgaagttc atctgcacct ccggcgagct gcccgtgccc
     3541 tggcccaccc tcgtgaccac cctgtcttac ggcgtgcagt gcttcgcccg ctaccccgac
     3601 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3661 accatctcct tcaaggacga cggcagctac aggacccgcg ccgaggtgaa gttcgagggc
     3721 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3781 ctggggcaca agctggagta caacatgaac gcgtgggacg cgtatatcac ggccgacaag
     3841 cagaagaacg gcatcaaagc gaacttcaag atcgagcaca acgtcaagga cggcggcgtg
     3901 cagctcgccg acgcgtacca gcagaacacc cccatcggcg acggctccgt gctgctgcct
     3961 gacaaccact acctgagctt ccagagcaag ctgttcaaag accccaacga gcagcgcgat
     4021 cacatggtcc tgctggagtt cgttaccgcc gccgggataa ctcccggcat ggacgagctg
     4081 tacaagtaaa gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc
     4141 ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc
     4201 tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4261 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     4321 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     4381 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4441 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4501 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4561 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4621 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     4681 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     4741 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     4801 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     4861 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     4921 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4981 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5041 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5101 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5161 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5221 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5281 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5341 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5401 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5461 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5521 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5581 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5641 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5701 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5761 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5821 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5881 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5941 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6001 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6061 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6121 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6181 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6241 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6301 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6361 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6421 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6481 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6541 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6601 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6661 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6721 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6781 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6841 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6901 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6961 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7021 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7081 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7141 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     7201 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7261 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7321 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     7381 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     7441 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7501 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     7561 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     7621 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     7681 ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac
     7741 aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata
     7801 gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa gggggg
//
