LOCUS       Exported                4762 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    L2963
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4762)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4762)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4762
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             10..30
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(73..101)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     918..938
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1960..1982)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2082..2101
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2179..2197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2265..2369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2370..3230
                     /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(2588..2607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3401..3989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3890..3909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4143..4160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4277..4298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4313..4343
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4351..4367
                     /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)."
     primer_bind     4356..4378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4375..4391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4375..4391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgactgctc caaagaagaa gcgtaaggta ccggtagaaa aaatgagtaa aggagaagaa
       61 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      121 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
      181 atttgcacta ctggaaaact acctgttcca tgggtaagtt taaacatata tatactaact
      241 aaccctgatt atttaaattt tcagccaaca cttgtcacta ctttctgtta tggtgttcaa
      301 tgcttctcga gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc
      361 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
      421 aagtttaaac agttcggtac taactaacca tacatattta aattttcagg tgctgaagtc
      481 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
      541 gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc
      601 atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt taaacatgat tttactaact
      661 aactaatctg atttaaattt tcagaacttc aaaattagac acaacattga agatggaagc
      721 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
      781 ccagacaacc attatctaag gtaagtttaa atgcatggtg accaacgttg ctagcgctac
      841 taactaactt gtactatcaa atttttcagc acacaatctg ccctttccaa agatcccaac
      901 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
      961 atggatgaac tatacaaata gcattcgtag aattgtaagt ttaaactttc tcatactaac
     1021 taacgcgctc tatttaaatt ttcagaattc caactgagcg ccggtcgcta ccattaccaa
     1081 cttgtctggt gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc
     1141 tactaactaa cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta
     1201 agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt
     1261 tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc
     1321 tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa
     1381 aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca caatgttctg
     1441 tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc
     1501 gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt
     1561 cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat
     1621 ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt
     1681 ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca
     1741 agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg
     1801 aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa
     1861 atgacagaat acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg
     1921 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1981 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2041 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     2101 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2161 catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2221 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2281 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2341 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2401 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2461 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2521 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2581 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2641 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2701 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2761 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2821 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2881 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2941 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3001 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3061 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3121 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3181 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3241 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3541 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3781 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4141 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4201 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4261 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     4321 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     4381 acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc
     4441 caattcttga gacttccact atcttctctg gcaccctcca ctacgcgtag gcgtctctcg
     4501 cttcgtgtat tcccgggaag ccgggttccc gtctctcccg ccgctgccgc tgccgcacac
     4561 agctttacac ctcgtagaat ccccaaagag gggcgtggct tgcgggtgcc aacatcctcc
     4621 tgccgaggaa gaagcaggca ctcatcactc gcatcatcaa cctcgggatt ggccaaagga
     4681 cccaaaggta tgtttcgaat gatactaaca taacatagaa cattttcagg aggacccttg
     4741 gagggtaccg agctcagaaa aa
//
