LOCUS       Exported                5112 bp ds-DNA     circular SYN 19-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5112)
  AUTHORS   emilianozamponi
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2022 from SnapGene Viewer 4.0.5
            http://www.snapgene.com
COMMENT     Sequenced 21-5-3
FEATURES             Location/Qualifiers
     source          1..5112
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    4..414
                     /label=Rtn2b-NTD136-271
     primer_bind     197..396
                     /label=Primer 2
     primer_bind     397..596
                     /label=Primer 3
     regulatory      445..454
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             451..1161
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      706..715
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    1283..1404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1411..1866)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1893..1997
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1999..2356
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2207..2342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2391..3185
                     /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"
     polyA_signal    3417..3464
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3793..4381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        4556..4859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4860..5063
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5109..196
                     /label=Primer 1
ORIGIN
        1 gctatgcctc tggaagacct gaggcttcgg ttggaccatc tgggctgggt ggcccgggga
       61 acgggatccg gggaggactc ttccaccagc agctccaccc cgctggaaga cgaagaaccc
      121 caagaaccca acagattgga gacaggagaa gctggggaag aactggacct acgactccga
      181 cttgctcagc cctcatcgcc cgaggtcttg actccccagc tcagtccggg ctctgggaca
      241 ccccaggccg gtactccgtc cccatcccga tcgcgagatt cgaactctgg gcccgaagag
      301 ccattgctgg aagaggaaga aaagcagtgg gggccactgg agcgagagcc agtaagggga
      361 cagtgcctcg atagcacgga ccaattagaa ttcacggtgg agccacgcct tctacgggta
      421 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg
      481 gcctctctcc cagcgacaca tgagttacac atctttggct ccatcaacgg tgtggacttt
      541 gacatggtgg gtcagggcac cggcaatcca aatgatggtt atgaggagtt aaacctgaag
      601 tccaccaagg gtgacctcca gttctccccc tggattctgg tccctcatat cgggtatggc
      661 ttccatcagt acctgcccta ccctgacggg atgtcgcctt tccaggccgc catggtagat
      721 ggctccggct accaagtcca tcgcacaatg cagtttgaag atggtgcctc ccttactgtt
      781 aactaccgct acacctacga gggaagccac atcaaaggag aggcccaggt gaaggggact
      841 ggtttccctg ctgacggtcc tgtgatgacc aactcgctga ccgctgcgga ctggtgcagg
      901 tcgaagaaga cttaccccaa cgacaaaacc atcatcagta cctttaagtg gagttacacc
      961 actggaaatg gcaagcgcta ccggagcact gcgcggacca cctacacctt tgccaagcca
     1021 atggcggcta actatctgaa gaaccagccg atgtacgtgt tccgtaagac ggagctcaag
     1081 cactccaaga ccgagctcaa cttcaaggag tggcaaaagg cctttaccga tgtgatgggc
     1141 atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt
     1201 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1261 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1321 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1381 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     1441 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1501 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1561 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1621 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1681 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1741 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1801 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1861 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1921 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1981 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2101 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2161 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2221 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2281 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2341 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     2401 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2461 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2521 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     2581 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2641 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2701 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2761 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2821 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2881 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     2941 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3001 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3061 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3121 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3181 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3241 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3301 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     3361 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     3421 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     3481 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3541 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3601 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     3661 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3721 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4441 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     4501 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4561 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4621 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4681 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4741 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4801 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     4861 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4921 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4981 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5041 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga
     5101 ctcagatctc ga
//
