LOCUS       Exported                8452 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 8452)
  AUTHORS   emilianozamponi
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2022 from SnapGene Viewer 4.0.5
            http://www.snapgene.com
COMMENT     Sequenced_21-5-4
FEATURES             Location/Qualifiers
     source          1..8452
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    7..1032
                     /label=TurboID
     CDS             10..51
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1048..1636
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(1519..1530)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1723..1956
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2028..2149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2189..2324
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2345..2363)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2373..2389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2531..2986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3012..3116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3117..3977
                     /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"
     rep_origin      4148..4736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5024..5045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5060..5090
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5098..5114
                     /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     5122..5138
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5159..5177
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5205..5431
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5432..5612
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5659..5784
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6277..6510
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7006..7122
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        7145..7448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7449..7652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      7742..7751
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7748..8452
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      8003..8012
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 aaggccatgg gcaagcccat ccccaacccc ctgctgggcc tggacagcac cgctagcaaa
       61 gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt ccatagtggc
      121 gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca cattcagact
      181 ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc tctgcccgag
      241 cctatcccgc tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca
      301 gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag
      361 agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag agggaggaaa
      421 tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct gaagcgggga
      481 ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc gctgcgaaag
      541 ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag
      601 ctggcaggca tcctggtgga gctggccgga ataacaggcg atgctgcaca gatcgtcatt
      661 ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg
      721 atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc tacgctgatc
      781 cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc atatctgcca
      841 cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat tggggacaaa
      901 gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct ggaacaggac
      961 ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga aaagctgcag
     1021 gtgtacaagt aaagcggccg cgtcgacaat caacctctgg attacaaaat ttgtgaaaga
     1081 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1141 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1201 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     1261 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     1321 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     1381 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     1441 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     1501 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     1561 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     1621 tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg
     1681 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     1741 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     1801 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1861 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1921 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     1981 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     2041 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2101 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     2161 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2221 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2281 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     2341 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     2401 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2461 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2521 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3661 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4261 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4801 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4861 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4921 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4981 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5041 ctcattaggc accccaggct ttccacttta tgcttccggc tcgtatgttg tgtggaattg
     5101 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     5161 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
     5221 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
     5281 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
     5341 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
     5401 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
     5461 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     5521 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     5581 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     5641 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     5701 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     5761 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     5821 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     5881 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     5941 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     6001 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     6061 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     6121 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     6181 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     6241 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     6301 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     6361 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     6421 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     6481 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     6541 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     6601 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     6661 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     6721 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     6781 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     6841 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     6901 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     6961 cagatccatt cgattagtga acggatctcg acggtatcgg ttaactttta aaagaaaagg
     7021 ggggattggg gggtatagtg caggggaaag aatagtagac ataatagcaa cagacataca
     7081 aactaaagaa ttacaaaaac aaattacaaa attcaaaatt ttatcgataa gcttgggagt
     7141 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     7201 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     7261 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     7321 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     7381 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     7441 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     7501 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     7561 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     7621 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     7681 gacgccatcc acgctgtttt gacctccata gaagacaccg actctagagg atccaccggt
     7741 cgccaccatg gtgagcaagg gcgaggagga taacatggcc tctctcccag cgacacatga
     7801 gttacacatc tttggctcca tcaacggtgt ggactttgac atggtgggtc agggcaccgg
     7861 caatccaaat gatggttatg aggagttaaa cctgaagtcc accaagggtg acctccagtt
     7921 ctccccctgg attctggtcc ctcatatcgg gtatggcttc catcagtacc tgccctaccc
     7981 tgacgggatg tcgcctttcc aggccgccat ggtagatggc tccggctacc aagtccatcg
     8041 cacaatgcag tttgaagatg gtgcctccct tactgttaac taccgctaca cctacgaggg
     8101 aagccacatc aaaggagagg cccaggtgaa ggggactggt ttccctgctg acggtcctgt
     8161 gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg aagaagactt accccaacga
     8221 caaaaccatc atcagtacct ttaagtggag ttacaccact ggaaatggca agcgctaccg
     8281 gagcactgcg cggaccacct acacctttgc caagccaatg gcggctaact atctgaagaa
     8341 ccagccgatg tacgtgttcc gtaagacgga gctcaagcac tccaagaccg agctcaactt
     8401 caaggagtgg caaaaggcct ttaccgatgt gatgggcatg gacgagctgt ac
//
