LOCUS       Exported                4693 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    L2604
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4693)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4693
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             307..327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(370..398)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1146..1166
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2133..2155)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2255..2274
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2352..2370)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2438..2542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2543..3403
                     /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(2761..2780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3574..4162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4063..4082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4316..4333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4450..4471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4486..4516
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4524..4540
                     /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     4529..4551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4548..4564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4548..4564
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagacact tttcagctac ctagatacat
       61 ggatatcccc gcctcccaat ccacccaccc agggaaaaag aagggctcgc cgaaaatcaa
      121 agttatctcc aggctcgcgc atcccaccga gcggttgact tctctccacc acttttcatt
      181 ttaaccctcg gggtacggga ttggccaaag gacccaaagg tatgtttcga atgatactaa
      241 cataacatag aacattttca ggaggaccct tcttggaggg taccgagctc agaaaaaatg
      301 actgctccaa agaagaagcg taaggtaccg gtagaaaaaa tgagtaaagg agaagaactt
      361 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      421 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
      481 tgcactactg gaaaactacc tgttccatgg gtaagtttaa acatatatat actaactaac
      541 cctgattatt taaattttca gccaacactt gtcactactt tctgttatgg tgttcaatgc
      601 ttctcgagat acccagatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa
      661 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtaag
      721 tttaaacagt tcggtactaa ctaaccatac atatttaaat tttcaggtgc tgaagtcaag
      781 tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat
      841 ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt atacatcatg
      901 gcagacaaac aaaagaatgg aatcaaagtt gtaagtttaa acatgatttt actaactaac
      961 taatctgatt taaattttca gaacttcaaa attagacaca acattgaaga tggaagcgtt
     1021 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     1081 gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac
     1141 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta
     1201 tacaaatagc attcgtagaa ttccaactga gcgccggtcg ctaccattac caacttgtct
     1261 ggtgtcaaaa ataatagggg ccgctgtcat cagagtaagt ttaaactgag ttctactaac
     1321 taacgagtaa tatttaaatt ttcagcatct cgcgcccgtg cctctgactt ctaagtccaa
     1381 ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta
     1441 ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca
     1501 atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt
     1561 gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac
     1621 ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca
     1681 aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt
     1741 ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga
     1801 atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt
     1861 cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg
     1921 atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga
     1981 gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag
     2041 aatacattcc caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt
     2101 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2161 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2221 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     2281 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2341 cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2401 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3061 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3721 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3961 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4201 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4321 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4381 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4441 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4501 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     4561 tgaccatgat tacgccaagc tgtaagttta aacatgatct tactaactaa ctattctcat
     4621 ttaaattttc agagcttaaa aatggctgaa atcactcaca acgatggata cgctaacaac
     4681 ttggaaatga aat
//
