LOCUS       Exported                9819 bp ds-DNA     circular SYN 10-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9819)
  AUTHORS   StevenLee
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 25, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9819
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             117..750
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    797..922
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1415..1648
                     /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    2164..2281
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2409..3587
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2640..3578
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3390..3409
                     /label=for
     primer_bind     3589..3629
                     /label=BD1 For
     misc_feature    3619..3975
                     /label=BD1
     primer_bind     3748..3772
                     /label=Primer 1
     CDS             3976..4656
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLE
                     FLTAA"
     misc_feature    4657..4758
                     /label=34-mer
     primer_bind     4657..4715
                     /label=34-mer F
     primer_bind     complement(4696..4758)
                     /label=34-mer R
     primer_bind     4739..4778
                     /label=ECFP 0914 F
     misc_feature    4759..5442
                     /label=ECFP
     primer_bind     complement(5427..5461)
                     /label=ECFP-H3-Rev
     regulatory      5437..5443
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    5443..5850
                     /label=H3
     primer_bind     5443..5481
                     /label=H3K14A-For
     primer_bind     5443..5466
                     /label=H3K9A-For
     CDS             5467..5469
                     /codon_start=1
                     /label=K9A
                     /translation="K"
     CDS             5482..5484
                     /codon_start=1
                     /label=K14A
                     /translation="K"
     primer_bind     complement(5834..5874)
                     /label=H3 Rev
     primer_bind     complement(5899..5920)
                     /label=rev
     misc_feature    5935..6508
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             6541..7140
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             7343..7576
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7733..7957
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      8000..8009
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        8053..8157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8158..9018
                     /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      9189..9777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
       61 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
      121 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
      181 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
      241 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
      301 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
      361 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
      421 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
      481 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
      541 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      601 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      661 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      721 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      781 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      841 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      901 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      961 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1021 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1081 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1141 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1201 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1261 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1321 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1381 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1441 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1501 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1561 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1621 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1681 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1741 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1801 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1861 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1921 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1981 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2041 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2101 gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag tattcatcca
     2161 caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2221 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2281 tcgggtttat tacagggaca gcagagatcc actttggtcg ataagctttg caaagatgga
     2341 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     2401 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2461 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     2521 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     2581 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     2641 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     2701 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     2761 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     2821 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     2881 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     2941 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     3001 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     3061 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     3121 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     3181 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     3241 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     3301 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     3361 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     3421 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     3481 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     3541 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attgatcctt
     3601 cgaactagtg ccaccatgtc caaccctaac aagcccaaga ggcagaccaa ccaactgcaa
     3661 tacctgctca gagtggtgct caagacacta tggaaacacc agtttgcatg gcctttccag
     3721 cagcctgtgg atgccgtcaa gctgaacctc cctgattact ataagatcat taaaacgcct
     3781 atggatatgg gaacaataaa gaagcgcttg gaaaacaact attactggaa tgctcaggaa
     3841 tgtatccagg acttcaacac tatgtttaca aattgttaca tctacaacaa gcctggagat
     3901 gacatagtct taatggcaga agctctggaa aagctcttct tgcaaaaaat aaatgagcta
     3961 cccacagaag aaaccatgtc taaaggtgaa gaattattca ctggtgttgt cccaattttg
     4021 gttgaattag atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt
     4081 gatgctacgt acggtaaatt gaccttaaaa ttactctgta ctactggtaa attgccagtt
     4141 ccatggccaa ccttagtcac tactttaggt tatggtgttc aatgttttgc tagataccca
     4201 gatcatatga aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa
     4261 agaactattt ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa
     4321 ggtgatacct tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac
     4381 attttaggtc acaaattgga atacaactat aactctcaca atgtttacat cactgctgac
     4441 aaacaaaaga atggtatcaa agctaacttc aaaattagac acaacattga agatggtggt
     4501 gttcaattag ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta
     4561 ccagacaacc attacttatc ctatcaatct gccttattca aagatccaaa cgaaaagaga
     4621 gatcacatgg tcttgttaga atttttgact gctgctggtt caacttctgg ctctgggaaa
     4681 ccaggaagcg gcgaagggtc caccaaggga agcaccagtg gttcaggtaa gcctggttct
     4741 ggtgaaggta gcactaaaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4801 atcctggtcg agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc
     4861 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4921 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc
     4981 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     5041 caggagcgta ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     5101 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     5161 ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc
     5221 gccgacaagc agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac
     5281 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     5341 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     5401 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgcacgaac caagcagact
     5461 gcccgcaaat ccaccggcgg taaggcacca cggaagcagc tggctaccaa agctgctaga
     5521 aagagtgctc ctgcaacagg gggagtgaaa aagcctcacc gatatcgccc tgggactgtg
     5581 gcccttaggg agatcagaag ataccaaaaa agcacagagt tgctgatccg gaagctgcca
     5641 tttcaacgac tggtgcggga gatagcccag gactttaaga ccgatcttag gttccaatcc
     5701 tcagctgtta tggcactcca ggaggccagt gaagcttacc tcgttgggtt gtttgaagac
     5761 accaacctgt gcgccatcca cgccaaaagg gtgacaatca tgccgaagga tattcagctc
     5821 gccagaagaa tcagagggga gagggcttag taggaattcg aagcttgagc tcgagatctg
     5881 agtccggtag cgctagttcg aaggatccgc atgcatctag ggcggccaat tccgcccctc
     5941 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     6001 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     6061 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     6121 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     6181 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     6241 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     6301 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     6361 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     6421 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     6481 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccacaaggag gcgaccttcc
     6541 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     6601 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     6661 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     6721 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     6781 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     6841 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     6901 cccgcgtggt tcctggccac cgtcggcgtt tcgcccgacc accagggcaa gggtctgggc
     6961 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7021 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7081 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     7141 taataggcgg ccgctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     7201 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     7261 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     7321 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     7381 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7441 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7501 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7561 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     7621 aagaaatgaa tatcagagag tgagaggcct tgacattata atagatttag caggaattga
     7681 actaggagtg gagcacacag gcaaagttct agagctcgct gatcagcctc gactgtgcct
     7741 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     7801 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     7861 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     7921 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     7981 tgggggcgcg cccctcgagg ccgccatggt catagctgtt tgacgtcagg tggcactttt
     8041 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     8101 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     8161 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     8221 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     8281 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     8341 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     8401 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     8461 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8521 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8581 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8641 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8701 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8761 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8821 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     8881 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     8941 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     9001 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     9061 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     9121 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     9181 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     9241 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     9301 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     9361 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9421 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9481 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9541 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     9601 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9781 gccagcaacg cggccttttt acggttcctg gccttttgc
//
