LOCUS       Exported                5256 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCR4-24XMS2SL-stable
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  AUTHORS   Bertrand E, Chartrand P, Schaefer M, Shenoy SM, Singer RH, Long RM
  TITLE     Localization of ASH1 mRNA particles in living yeast.
  JOURNAL   Mol Cell. 1998 Oct . 2(4):437-45.
  PUBMED    9809065
REFERENCE   2  (bases 1 to 5256)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5256
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(35..53)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(74..90)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(74..90)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(87..109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    98..114
                     /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(122..152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(305..322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(476..1064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(556..575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1372..1750
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(1406..1425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2296..3090)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2407..2426)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3017..3036
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3439..78)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSELTLTKGTSPAGLNEFALRSDEPWNTGATRDRRRLKI
                     RPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPW
                     NTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARA
                     FLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRR
                     LKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSD
                     EPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTAD
                     ARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRD
                     RRRLKIRPGGTADARAFLAISTVGSDEPWNTGATRDRRRLKIRPGGTADARAFLAISTV
                     GSKGEFAAAKFNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLYVRQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3541..3560)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3864..3886
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3878..3895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3879..3895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3902..3921
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3902..3920
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagggcgaat tcgtttaaac ctgcaggact agtcccttta gtgagggtta attctgagct
       61 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      121 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      181 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      241 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      301 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      361 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      901 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1141 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1201 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1261 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1321 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1381 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1441 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1501 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1561 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1621 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1681 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1741 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1801 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gccttccggc
     1861 tggctggttt attgctgata aatctggagc cggtgagcgt ggtctcgcgg tatcattgca
     1921 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1981 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2041 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2101 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgagcgg atacatattt
     2161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2221 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     2281 gtaagcgtta ataattcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     2341 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     2401 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     2461 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     2521 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa
     2581 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     2641 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     2701 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     2761 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     2821 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     2881 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
     2941 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     3001 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     3061 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     3121 agagcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     3181 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt
     3241 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
     3301 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     3361 gcaccgtttc tgcggactgg ctttctacgt gaaaaggatc taggtgaaga tcctttttga
     3421 taatctcatg cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc
     3481 attttcgcgg tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc
     3541 catatcggtg gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc
     3601 acgggagact ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc
     3661 cggcgtgtca ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt
     3721 ataggtgtaa accttaaact gccgtacgta taggctgcgc aactgttggg aagggcgatc
     3781 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     3841 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     3901 gtaatacgac tcactatagg gcgaattgaa tttagcggcc gcgaattcgc ccttggatcc
     3961 tacggtactt attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa
     4021 acgacgacga tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat
     4081 tgccaagaaa gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc
     4141 acgcgtcgct ccagtattcc agggttcatc agatcctacg gtacttattg ccaagaaagc
     4201 acgagcatca gccgtgcctc caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc
     4261 agtattccag ggttcatcag atcctacggt acttattgcc aagaaagcac gagcatcagc
     4321 cgtgcctcca ggtcgaatct tcaaacgacg acgatcacgc gtcgctccag tattccaggg
     4381 ttcatcagat cctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg
     4441 tcgaatcttc aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc
     4501 tacggtactt attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa
     4561 acgacgacga tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat
     4621 tgccaagaaa gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc
     4681 acgcgtcgct ccagtattcc agggttcatc agatcctacg gtacttattg ccaagaaagc
     4741 acgagcatca gccgtgcctc caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc
     4801 agtattccag ggttcatcag atcctacggt acttattgcc aagaaagcac gagcatcagc
     4861 cgtgcctcca ggtcgaatct tcaaacgacg acgatcacgc gtcgctccag tattccaggg
     4921 ttcatcagat cctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg
     4981 tcgaatcttc aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc
     5041 tacggtactt attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa
     5101 acgacgacga tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat
     5161 tgccaagaaa gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc
     5221 acgcgtcgct ccagtattcc agggttcatc agatct
//
