LOCUS       sequence-133147-        7938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7938)
  TITLE     Littman Lab Plasmids
REFERENCE   2  (bases 1 to 7938)
  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"
COMMENT     Sequence Label: MIGR-RORgt
FEATURES             Location/Qualifiers
     source          1..7938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2365..2387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2515..2534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2734..3250
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3314..3655
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3722..4138
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4068..4090
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4106..4122
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5664..6231
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5681..6233
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5825..5842)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6052..6071
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6245..6247,6248..6250,6251..6964)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6290..6311)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6551..6570)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6898..6919
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             7066..7580
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7742..7758
                     /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(7766..7772,7773..7790,7791..7796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7811..7832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa
     2401 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     2461 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     2521 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     2581 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     2641 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     2701 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     2761 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     2821 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     2881 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     2941 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3001 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3061 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3121 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa
     3181 tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg
     3241 gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc
     3301 cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt
     3361 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat
     3421 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     3481 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     3541 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     3601 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     3661 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     3721 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     3781 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     3841 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     3901 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     3961 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     4021 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     4081 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     4141 aattagatcc accatgagaa cacaaattga agtgatccct tgcaagatct gtggggacaa
     4201 gtcatctgga atccactacg gggttatcac ctgtgagggg tgcaagggct tcttccgccg
     4261 cagccagcag tgtaatgtgg cctactcctg cacgcgtcag cagaactgcc ccattgaccg
     4321 aaccagccgc aaccgatgcc agcattgccg cctgcagaag tgcctggctc tgggcatgtc
     4381 ccgagatgct gtcaagtttg gccgaatgtc caagaagcag agggacagtc tacatgcaga
     4441 agtgcagaaa caactgcaac agcagcagca acaggaacaa gtggccaaga ctcctccagc
     4501 tgggagccgc ggagcagaca cacttacata cactttaggg ctctcagatg ggcagctacc
     4561 actgggcgcc tcacctgacc tacccgaggc ctctgcttgt ccccctggcc tcctgagagc
     4621 ctcaggctct ggcccaccat attccaatac cttggccaaa acagaggtcc agggggcctc
     4681 ctgccacctt gagtatagtc cagaacgagg caaagctgaa ggcagagaca gcatctatag
     4741 cactgacggc caacttactc ttggaagatg tggacttcgt tttgaggaaa ccaggcatcc
     4801 tgaacttggg gaaccagaac agggtccaga cagccactgc attcccagtt tctgcagtgc
     4861 cccagaggta ccatatgcct ctctgacaga catagagtac ctggtacaga atgtctgcaa
     4921 gtccttccga gagacatgcc agctgcgact ggaggacctt ctacggcagc gcaccaacct
     4981 cttttcacgg gaggaggtga ccagctacca gaggaagtca atgtgggaga tgtgggagcg
     5041 ctgtgcccac cacctcactg aggccattca gtatgtggtg gagtttgcca agcggctttc
     5101 aggcttcatg gagctctgcc agaatgacca gatcatacta ctgaaagcag gagcaatgga
     5161 agtcgtccta gtcagaatgt gcagggccta caatgccaac aaccacacag tcttttttga
     5221 aggcaaatac ggtggtgtgg agctgtttcg agccttgggc tgcagcgagc tcatcagctc
     5281 catatttgac ttttcccact tcctcagcgc cctgtgtttt tctgaggatg agattgccct
     5341 ctacacggcc ctggttctca tcaatgccaa ccgtcctggg ctccaagaga agaggagagt
     5401 ggaacatctg caatacaatt tggaactggc tttccatcat catctctgca agactcatcg
     5461 acaaggcctc ctagccaagc tgccacccaa aggaaaactc cggagcctgt gcagccaaca
     5521 tgtggaaaag ctgcagatct tccagcacct ccaccccatc gtggtccaag ccgccttccc
     5581 tccactctat aaggaactct tcagcactga tgttgaatcc cctgaggggc tgtcaaagtg
     5641 actcgagaaa aaaaagggcg aattccgccc ccccccccta acgttactgg ccgaagccgc
     5701 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     5761 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     5821 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     5881 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     5941 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     6001 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     6061 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     6121 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     6181 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     6241 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     6301 ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac
     6361 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     6421 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     6481 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     6541 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     6601 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     6661 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     6721 gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     6781 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     6841 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     6901 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     6961 gtaatgaatt aattaagaat tatcaagctt atcgataccg tcgacctgca gccaagctta
     7021 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     7081 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac
     7141 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa
     7201 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     7261 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     7321 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     7381 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     7441 cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     7501 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     7561 cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg
     7621 gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact
     7681 ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa
     7741 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7801 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7861 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7921 gtttgcgtat tgggcgct
//
