LOCUS       Exported                4913 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    L2603
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4913)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4913)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4913
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             527..547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(590..618)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1366..1386
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2353..2375)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2475..2494
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2572..2590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2658..2762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2763..3623
                     /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(2981..3000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3794..4382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4283..4302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4536..4553
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4670..4691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4706..4736
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4744..4760
                     /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     4749..4771
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4768..4784
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4768..4784
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagaggat ccccggggat tggccaaagg
       61 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      121 gcttggctag caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca
      181 gcatccgcac tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc
      241 gttgtagggg gcggagcgat aggtcctata ggttttggta tatcatcatt cattcattca
      301 ttggtacatt catttaccca ccttcctctt tctgagcttc tctggagttc tgtgcttcct
      361 ttttccctta tctttatact gtaattttta actttcaggc attgattgga tccccgggat
      421 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      481 gaggaccctt gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      541 taaggtaccg gtagaaaaaa tgagtaaagg agaagaactt ttcactggag ttgtcccaat
      601 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      661 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      721 tgttccatgg gtaagtttaa acatatatat actaactaac cctgattatt taaattttca
      781 gccaacactt gtcactactt tctgttatgg tgttcaatgc ttctcgagat acccagatca
      841 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
      901 tatatttttc aaagatgacg ggaactacaa gacacgtaag tttaaacagt tcggtactaa
      961 ctaaccatac atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt
     1021 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     1081 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     1141 aatcaaagtt gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca
     1201 gaacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     1261 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     1321 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     1381 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagc attcgtagaa
     1441 ttccaactga gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg
     1501 ccgctgtcat cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt
     1561 ttcagcatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac
     1621 atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta
     1681 atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa
     1741 aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata
     1801 ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct
     1861 gataaatttt tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta
     1921 cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca
     1981 aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag
     2041 tttatgaaat taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat
     2101 ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag
     2161 atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa
     2221 agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca
     2281 aacataactg tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga
     2341 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2401 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2461 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     2521 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga
     2581 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2641 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2701 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2761 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2821 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2881 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2941 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3001 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3061 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3121 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3181 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3241 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3301 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3361 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3421 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3481 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3541 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3601 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3661 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3721 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3781 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3841 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3901 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3961 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4021 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4081 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4141 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     4201 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4261 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4321 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4381 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4441 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4501 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4561 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4621 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4681 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4741 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     4801 tgtaagttta aacatgatct tactaactaa ctattctcat ttaaattttc agagcttaaa
     4861 aatggctgaa atcactcaca acgatggata cgctaacaac ttggaaatga aat
//
