LOCUS       Exported                9614 bp DNA     circular SYN 30-JUN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9614)
  AUTHORS   Mario Escobar
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9614
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     CDS             complement(344..355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             533..766
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    798..1022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1068..1496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1510..1839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        1887..1934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1953..2327
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2457..2578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2651..2667
                     /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(2675..2705)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2720..2741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3029..3617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3788..4648)
                     /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"
     promoter        complement(4649..4753)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5019..5398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5399..5602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             5616..5796
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5843..5968
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6461..6694
                     /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    7221..7338
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        7595..7806
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     CDS             7831..8550
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        8557..9067
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     regulatory      9068..9077
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9078..9476
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    join(9487..9614,1..461)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
ORIGIN
        1 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
       61 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
      121 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
      181 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
      241 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
      301 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
      361 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
      421 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
      481 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
      541 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
      601 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      661 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      721 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
      781 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
      841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
      901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
      961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1141 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1201 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1261 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1441 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1501 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1561 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1681 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1741 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     1801 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     1861 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     1921 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     1981 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2041 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2101 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2161 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2221 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     2281 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     2341 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2401 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2461 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2521 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2581 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     2641 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2701 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4741 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     4801 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4861 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     4921 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     4981 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     5041 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5101 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5161 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5221 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     5281 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5341 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     5401 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     5461 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     5521 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     5581 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     5641 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5701 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5761 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5821 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     5881 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     5941 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6001 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6061 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6121 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6181 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6241 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6301 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     6361 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     6421 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     6481 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     6541 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     6601 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     6661 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     6721 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     6781 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     6841 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     6901 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     6961 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     7021 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     7081 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     7141 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     7201 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     7261 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     7321 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     7381 aaggtaccga tatcactagt gttccattaa ctgcgctaaa aataattttt aaatcttttt
     7441 tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca agttaaaata
     7501 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct
     7561 agcgtcccct ccaccccaca gtctcgaggg gcgagggcag agcgcacatc gcccacagtc
     7621 cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg
     7681 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     7741 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     7801 acacaggacc ggttctagag cgctgccacc atggtgagca agggcgagga gctgttcacc
     7861 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     7921 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     7981 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     8041 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     8101 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     8161 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     8221 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     8281 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     8341 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     8401 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     8461 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     8521 actctcggca tggacgagct gtacaagtga ggatccgggg ttggggttgc gccttttcca
     8581 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     8641 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     8701 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     8761 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     8821 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     8881 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     8941 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     9001 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     9061 tccccaggcc accatggatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     9121 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     9181 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     9241 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     9301 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     9361 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     9421 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaatcga
     9481 ttcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     9541 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     9601 tcccgtatgg cttt
//
