LOCUS       sequence-258469-        6282 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 6282)
  AUTHORS   Patel MR, Loo YM, Horner SM, Gale M Jr, Malik HS
  TITLE     Convergent evolution of escape from hepaciviral antagonism in 
            primates.
  JOURNAL   PLoS Biol. 2012;10(3):e1001282. doi: 10.1371/journal.pbio.1001282. 
            Epub 2012 Mar 13.
  PUBMED    22427742
REFERENCE   2  (bases 1 to 6282)
  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"; journalName: "PLoS 
            Biol."; date: "2012"; volume: "10(3):e1001282. doi"; pages: " 
            10.1371/journal.pbio.1001282. Epub 2012 Mar 13"
COMMENT     Sequence Label: pMP31-1
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2623..3245
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2822..2841)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3269..3289)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3274..3603
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3454..3589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3641..3660)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3642..3660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3674..3690)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3674..3690)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3687..3709)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3698..3714
                     /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(3722..3728,3729..3746,3747..3752))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3767..3788
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3905..3922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4076..4664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4156..4175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4835..5626,5627..5695))
                     /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     5458..5477
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5696..5800)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5827..6255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5914..5933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6124..6145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tatgccgttt gctgaagaca agacctataa gtatatctgc cgcaatttca gcaatttttg
     1021 caatgtggat gttgtagaga ttctgcctta cctgccctgc ctcacagcaa gagaccagga
     1081 tcgactgcgg gccacctgca cactctcagg gaaccgggac accctctggc atctcttcaa
     1141 tacccttcag cggcggcccg gctgggtgga gtacttcatt gcggcactga ggggctgtga
     1201 gctagttgat ctcgcggacg aagtggcctc tgtctaccag agctaccagc ctcggacctc
     1261 ggaccgtccc ccagacccac tggagccacc gtcacttcct gctgagaggc cagggccccc
     1321 cacacctgct gcggcccaca gcatccccta caacagctgc agagagaagg agccaagtta
     1381 ccccatgcct gtccaggaga cccaggcgcc agagtcccca ggagagaatt cagagcaagc
     1441 cctgcagacg ctcagcccca gagccatccc aaggaatcca gatggtggcc ccctggagtc
     1501 ctcctctgac ctggcagccc tcagccctct gacctccagc gggcatcagg agcaggacac
     1561 agaactgggc agtacccaca cagcaggtgc gacctccagc ctcacaccat cccgtgggcc
     1621 tgtgtctcca tctgtctcct tccagcccct ggcccgttcc acccccaggg caagccgctt
     1681 gcctggaccc acagggtcag ttgtatctac tggcacctcc ttctcctcct catcccctgg
     1741 cttggcctct gcaggggctg cagagggtaa acagggtgca gagagtgacc aggccgagcc
     1801 tatcatctgc tccagtgggg cagaggcacc tgccaactct ctgccctcca aagtgcctac
     1861 caccttgatg cctgtgaaca cagtggccct gaaagtgcct gccaacccag catctgtcag
     1921 cacagtgccc tccaagttgc caactagctc aaagccccct ggtgcagtgc cttctaatgc
     1981 gctcaccaat ccagcaccat ccaaattgcc catcaactca acccgtgctg gcatggtgcc
     2041 atccaaagtg cctactagca tggtgctcac caaggtgtct gccagcacag tccccactga
     2101 cgggagcagc agaaatgagg agaccccagc agctccaaca cccgccggcg ccactggagg
     2161 cagctcagcc tggctagaca gcagctctga gaataggggc cttgggtcgg agctgagtaa
     2221 gcctggcgtg ctggcatccc aggtagacag cccgttctcg ggctgcttcg aggatcttgc
     2281 catcagtgcc agcacctcct tgggcatggg gccctgccat ggcccagagg agaatgagta
     2341 taagtccgag ggcacctttg ggatccacgt ggctgagaac cccagcatcc agctcctgga
     2401 gggcaaccct gggccacctg cggacccgga tggcggcccc aggccacaag ccgaccggaa
     2461 gttccaggag agggaggtgc catgccacag gccctcacct ggggctctgt ggctccaggt
     2521 ggctgtgaca ggggtgctgg tagtcacact cctggtggtg ctgtaccggc ggcgtctgca
     2581 ctagagatct gatatcggta ccagtcgact ctagaggatc ccgggtggca tccctgtgac
     2641 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2701 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2761 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2821 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2881 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2941 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3001 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3061 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac
     3121 cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat
     3181 ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt
     3241 gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3421 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3481 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3541 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3601 aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt ccctatagtg agtcgtatta
     3661 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3721 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3781 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3841 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3901 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3961 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4021 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4081 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4141 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4201 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4261 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4321 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4381 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4441 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4501 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4561 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4621 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4681 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4741 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4801 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4861 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4921 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4981 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5041 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5101 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5161 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5221 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5281 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5341 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5401 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5461 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5521 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5581 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5641 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5701 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5761 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5821 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5881 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5941 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6001 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     6061 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6121 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6181 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6241 aatttaacgc gaattttaac aaaatattaa cgcttacaat tt
//
