LOCUS       sequence-275497-        6492 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 6492)
  TITLE     Viral vectors.
REFERENCE   2  (bases 1 to 6492)
  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: pMIG
FEATURES             Location/Qualifiers
     source          1..6492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(35..451)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(51..67)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(83..105)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    518..859
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             923..1439
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1639..1658)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1786..1808)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2002..2021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2121..2143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2181..2199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2267..2371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2372..2440,2441..3232)
                     /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(2590..2609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3403..3991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3892..3911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4145..4162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4279..4300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4315..4320,4321..4338,4339..4345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4353..4369
                     /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)."
     LTR             4531..5045
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(5195..5908,5909..5911,5912..5914))
                     /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(5240..5261)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5589..5608
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5848..5869
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    5926..6478
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6088..6107)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6317..6334
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 ggcggaattc gttaacctcg agagatctaa ttccggcgcc tagagaagga gtgagggctg
       61 gataaaggga ggatcgaggc ggggtcgaac gaggaggttc aagggggaga gacggggcgg
      121 atggaggaag aggaggcgga ggcttagggt gtacaaaggg cttgacccag ggaggggggt
      181 caaaagccaa ggcttcccag gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc
      241 caggtgaaaa gaccttgatc ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc
      301 gcggccatcc gacgttaaag gttggccatt ctgcagagca gaaggtaacc caacgtctct
      361 tcttgacatc taccgactgg ttgtgagcga tccgctcgac atctttccag tgacctaagg
      421 tcaaacttaa gggagtggta acagtctggc cctaattttc agacaaatac agaaacacag
      481 tcagacagag acaacacaga acgatgctgc agcgctgcag cagacaagac gcgcggcttc
      541 ggttccaaac cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg ttctcgtctc
      601 ctaccagaac cacatatcct gacggggtcg gattccacat cgactccctt cctcaggtcg
      661 ggccacaaaa acggccccca aagtccctgg gacgtctccc agggttgcgg ccgggtgttc
      721 agaactcgtc agttccacca cgggtccgcc agatacagag ctagttagct aactagtaca
      781 gacgcaggcg caaacattag atgccggcac aaacacgcac aaagacagag acagacacga
      841 aacgaccgct ggccagctta cctcccggcg ggggggtcgg tggtccctgg gcaggggtct
      901 ccaaatctcg gtggaacctc caaatgaaag acccccgagg tgggcagtca atcaatctga
      961 ggagaccctc ccaaggatca gcgagtccac gattcggatg caaacagcaa gaggctttat
     1021 tgggaatacg ggtacccggg cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg
     1081 ttgtgggctc ttttattgag ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa
     1141 ctgattggtt agttcaaata aggcacaggg tcatttcagg tccttggggc accctggaaa
     1201 catctgatgg ttctctagaa actgctgagg gcgggaccgc atctggggac catctgttct
     1261 tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc catattctgc
     1321 tgtctctctg ttcctaacct tgatctgaac ttctctattc tcagttatgt attttccatg
     1381 ccttgcaaaa tggcgttact taagctagct tgccaaacct acaggtgggg tctttcattc
     1441 cccccttttt ctggagacta aataaaatct tttattttat ctatggctcg tactctatag
     1501 gcttcagctg gtgatattgt tgagtcaaaa ctagagcctg gaccactgat atcctgtctt
     1561 taacaaattg gactaatcgc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     1621 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     1681 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     1741 gactgttggg cgccatctcc ttgcatgcac cattccttgc gccgtcgttt tacaacgtcg
     1801 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1861 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1921 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1981 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     2041 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2101 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2161 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2221 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2341 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2461 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2521 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2641 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2761 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2941 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3241 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3301 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3541 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4021 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4081 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4141 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     4201 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4261 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     4321 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacaattc
     4381 tccccagctc tttctgcgct gtccataatg aactatttca ggagtattgg agtatgtgga
     4441 ttcaaaacag tggctaattg agtcaggatt acttaatatt cgactcctac cctcagaatg
     4501 ttgttctcca acttcaagaa actgttaccc atgaaagacc cccgctgacg ggtagtcaat
     4561 cactcagagg agaccctccc aaggaacagc gagaccacaa gtcggatgca actgcaagag
     4621 ggtttattgg atacacgggt acccgggcga cgcagtctat cggaggactg gcgcgccgag
     4681 tgaggggttg tgggctcttt tattgagctc ggggagcaga agcgcgcgaa cagaagcgag
     4741 aagcgaactg attggttagt tcaaataagg cacagggtca tttcaggtcc ttggggcacc
     4801 ctggaaacat ctgatggttc tctagaaact gctgagggcg ggaccgcatc tggggaccat
     4861 ctgttcttgg ccctgagccg gggcaggaac tgcttaccac agatatcctg tttggcccat
     4921 attctgctgt ctctctgttc ctaaccttga tctgaacttc tctattctca gttatgtatt
     4981 ttccatgcct tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct
     5041 ttcattcccc cctttttctg gagactaaat aaaatctttt attttatcga taagcttggc
     5101 tgcaggtcga ctctagaaag tgtctcatgc cggaggccat taaagacagg ccttactggc
     5161 tgaatagaag cggtgataat tcttaattaa ttcattactt gtacagctcg tccatgccga
     5221 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     5281 ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
     5341 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     5401 tgttgtggcg gatcttgaag ttcgccttga tgccgttctt ctgcttgtcg gccatgatat
     5461 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     5521 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     5581 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     5641 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     5701 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     5761 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     5821 cagacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     5881 ccccggtgaa cagctcctcg cccttgctca ccatggttgt ggccatatta tcatcgtgtt
     5941 tttcaaagga aaaccacgtc cccgtggttc ggggggccta gacgtttttt taacctcgac
     6001 taaacacatg taaagcatgt gcaccgaggc cccagatcag atcccataca atggggtacc
     6061 ttctgggcat ccttcagccc cttgttgaat acgcttgagg agagccattt gactctttcc
     6121 acaactatcc aactcacaac gtggcactgg ggttgtgccg cctttgcagg tgtatcttat
     6181 acacgtggct tttggccgca gaggcacctg tcgccaggtg gggggttccg ctgcctgcaa
     6241 agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca gaggaactgc ttccttcacg
     6301 acattcaaca gaccttgcat tcctttggcg agaggggaaa gacccctagg aatgctcgtc
     6361 aagaagacag ggccaggttt ccgggccctc acattgccaa aagacggcaa tatggtggaa
     6421 aataacatat agacaaacgc acaccggcct tattccaagc ggcttcggcc agtaacgtta
     6481 gggggggggg ag
//
