LOCUS       Exported                7542 bp ds-DNA     circular SYN 29-JUN-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7542)
  AUTHORS   Brandon Cisneros
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 29, 2020 from SnapGene Viewer 5.1.4.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7542
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          5813..5831
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          6939..6951
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(109..213)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     CDS             214..1074
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:1: 5552 .. 6343 /#ccffcc
                     2: 6344 .. 6412 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1245..1833
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2017..2020
                     /label=tata
                     /note="tata"
     misc_feature    2021..2245
                     /label=3'-ITR
                     /note="3'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
     misc_feature    2246..2287
                     /label=3'-proxITR
                     /note="3'-proxITR"
                     /note="modified region proximal to 3'-ITR - contains CTCF 
                     binding sites"
     misc_feature    2298..2598
                     /label=rBG-pA
                     /note="rBG-pA"
                     /note="rabbit beta-globin polyadenylation signal"
     polyA_signal    2396..2451
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(2616..3215)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             complement(3216..3272)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(3282..3983)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein"
                     /label=dTomato (dimeric DsRed)
                     /note="dTomato"
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     promoter        3997..4608
                     /note="RPBSA"
                     /note="constitutive promoter"
     exon            3997..4006
                     /note="RPL41 exon2"
     intron          4007..4121
                     /note="RPL41 intron1"
     exon            4122..4174
                     /note="RPL41 exon1"
     promoter        4616..5797
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human 
                     elongationfactor EF-1-alpha"
     intron          complement(4846..5788)
                     /note="EF1a Intron1"
     exon            complement(5789..5807)
                     /note="EF1a Exon2"
     misc_feature    5813..5825
                     /label=5'-SfiI
                     /note="5'-SfiI"
                     /note="GGCCTCTGAGGCC"
     CDS             5829..6938
                     /codon_start=1
                     /product="caffeine synthase 1 [Camellia sinensis] "
                     /label=TCS1
                     /protein_id="BAB12278.1 "
                     /translation="MELATAGKVNEVLFMNRGEGESSYAQNSSFTQQVASMAQPALENA
                     VETLFSRDFHLQALNAADLGCAAGPNTFAVISTIKRMMEKKCRELNCQTLELQVYLNDL
                     FGNDFNTLFKGLSSEVIGNKCEEVPCYVMGVPGSFHGRLFPRNSLHLVHSSYSVHWLTQ
                     APKGLTSREGLALNKGKIYISKTSPPVVREAYLSQFHEDFTMFLNARSQEVVPNGCMVL
                     ILRGRQCSDPSDMQSCFTWELLAMAIAELVSQGLIDEDKLDTFNIPSYFASLEEVKDIV
                     ERDGSFTIDHIEGFDLDSVEMQENDKWVRGEKFTKVVRAFTEPIISNQFGPEIMDKLYD
                     KFTHIVVSDLEAKLPKTTSIILVLSKIDG"
     misc_feature    6939..6951
                     /label=3'-SfiI
                     /note="3'-SfiI"
                     /note="GGCCTGTCAGGCC"
     polyA_signal    complement(6952..7249)
                     /label=bGH-pA
                     /note="bGH-pA"
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7260..7301
                     /label=5'-proxITR
                     /note="5'-proxITR"
                     /note="modified region proximal to 5'-ITR - contains CTCF 
                     binding sites"
     misc_feature    7307..7531
                     /label=5'-ITR
                     /note="5'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
     misc_feature    7532..7535
                     /label=tata
                     /note="tata"
ORIGIN
        1 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
       61 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
      121 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      181 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      241 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      301 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      361 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      421 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      481 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      541 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      601 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      661 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      721 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      781 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      841 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      901 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      961 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1021 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1081 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1921 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1981 gagcgcagcg agtcagtgag cgaggaagcg gatccctata cagttgaagt cggaagttta
     2041 catacacctt agccaaatac atttaaactc actttttcac aattcctgac atttaatcct
     2101 agtaaaaatt ccctgtctta ggtcagttag gatcaccact ttattttaag aatgtgaaat
     2161 atcagaataa tagtagagag aatgattcat ttcagctttt atttctttca tcacattccc
     2221 agtgggtcag aagtttacat acactcgact cctctgcaga atgcggcgat gtttcggtaa
     2281 ggggtcccga gtagctagtt catggcagcc agcatatggc atatgttgcc aaactctaaa
     2341 ccaaatactc attctgatgt tttaaatgat ttgccctccc atatgtcctt ccgagtgaga
     2401 gacacaaaaa attccaacac actattgcaa tgaaaataaa tttcctttat tagccagaag
     2461 tcagatgctc aaggggcttc atgatgtccc cataattttt ggcagaggga aaaagatctc
     2521 agtggtattt gtgagccagg gcattggcca caccagccac caccttctga taggcagcct
     2581 gcacctgagg agtgaattca cgacaggcct tcgaatcagg caccgggctt gcgggtcatg
     2641 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     2701 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
     2761 tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     2821 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     2881 aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     2941 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     3001 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     3061 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     3121 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     3181 gtggcgaggc gcaccgtggg cttgtactcg gtcatggggc caggattctc ctcgacgtca
     3241 ccagcctgct tcagcaggct gaagttagta gctccactgc ccttgtacag ctcgtccatg
     3301 ccgtacagga acaggtggtg gcggccctcg gagcgctcgt actgttccac gatggtgtag
     3361 tcctcgttgt gggaggtgat gtccagcttg gtgtccacgt agtagtagcc gggcagttgc
     3421 acgggcttct tggccatgta gatggtcttg aactccacca ggtagtggcc gccgtccttc
     3481 agcttcaggg cctggtggat ctcgcccttc agcacgccgt cgcgggggta caggcgctcg
     3541 gtggaggcct cccagcccat tgtcttcttc tgcattacgg ggccgtcggg ggggaagttg
     3601 gtgccgcgca tcttcacctt gtagatcagc gtgccgtcct gcagggagga gtcctgggtc
     3661 acggtcacca gaccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg
     3721 aaggacagct tcttgtaatc ggggatgtcg gcggggtgct tcacgtacgc cttggagccg
     3781 tacatgaact ggggggacag gatgtcccag gcgaagggca gggggccgcc cttggtcacc
     3841 ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg
     3901 aactcatggc cgttcatgga gccctccatg cgcaccttga agcgcatgaa ctctttgatg
     3961 acctcctcgc ccttgctcaa catggtggcc tcaggtgcag aggtttctac agggaaagag
     4021 agaaaggcta cagttagttt gctgtgatga tgctaagtgc cgaggtcaga tgtctatgga
     4081 ttgtacacgg aatcccaacg caggaaccaa aaaacactca cccaaaaaaa tggcgctaag
     4141 gccgagagaa aggcgaagtt ccgtctacgg ctatatacct tgaaacaggg gcggcagaag
     4201 tgacgcaacg cggttgcgcg gacggcttcc tgtgttttca ttggtcgaag cgttctgcgc
     4261 gtgcgcagag attccccagg gaaggcgggc cctcttccga atgtccaatc cggacattct
     4321 tccggttgga cctccgcggt cagcgctgtg cccggcgaga gatcacgtgg ggcgcggagg
     4381 cggtgctgct ggggcacggc cgtccagcct cggcggccat atttttgagg ggctgttcat
     4441 ctcgttcaca cgctctgtcc gccatgtttg tgagtggaag cgccattacc ccttcaagcg
     4501 actgaaggct gcagggcctc tggtggcccg catggggaga ccagacccgc caggcccgcc
     4561 tttccgcact cagtccgggc ttactttatt ttgtgagaca gggtctcgcc tagaggctcc
     4621 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4681 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     4741 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     4801 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
     4861 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
     4921 acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga
     4981 gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcttggcc
     5041 tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt
     5101 cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca
     5161 agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc
     5221 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
     5281 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
     5341 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
     5401 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
     5461 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
     5521 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
     5581 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
     5641 gtttccccat actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
     5701 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
     5761 ggttcaaagt ttttttcttc catttaaggt gtcgtgaaaa ctaccccaag ctggcctctg
     5821 aggccaccat ggagctagct actgcgggga aggtgaacga agtgttgttc atgaacagag
     5881 gagaaggaga aagtagttat gcacaaaact cttctttcac gcaacaagtg gcctcaatgg
     5941 cacagccagc gctagaaaat gcagttgaaa ctctcttctc cagagatttc caccttcaag
     6001 ctcttaacgc agcggacttg ggttgtgcag cgggtccaaa cacattcgca gtgatttcta
     6061 cgatcaagag aatgatggaa aagaaatgca gggaattgaa ttgccaaaca ctggaacttc
     6121 aggtttactt gaatgatctt tttggaaatg atttcaatac cctcttcaaa ggcctgtcgt
     6181 ctgaggttat tggtaacaaa tgtgaggaag ttccgtgtta tgtgatggga gtaccggggt
     6241 ctttccatgg ccggcttttt cctcgtaaca gcttacattt agttcattcc tcttacagtg
     6301 ttcattggct tactcaggca ccaaaaggac tcacaagcag agaaggcttg gcattaaaca
     6361 aggggaagat ttacatatca aagacaagcc ctcctgttgt aagagaagcc tacttatctc
     6421 aatttcatga agatttcaca atgtttctca atgctagatc ccaagaggtg gttccaaatg
     6481 gttgtatggt gttgatactt cgtggtaggc aatgttctga tccttcagac atgcagagct
     6541 gctttacttg ggaactatta gctatggcca ttgctgaatt ggtttcacag ggattgatag
     6601 atgaagataa attagacacc ttcaatatac ccagctattt tgcatcactt gaggaagtga
     6661 aagatatagt ggagagggac ggatcattca caattgatca tatagagggg tttgatcttg
     6721 atagcgtaga aatgcaggag aatgataaat gggttagagg ggaaaagttt accaaggttg
     6781 tcagggcctt cacagagcct ataatttcaa accagtttgg acctgaaatc atggacaaac
     6841 tatatgacaa attcactcac attgtagttt cagatttgga agcaaagcta ccgaagacca
     6901 caagtatcat cctagtgctt tccaagattg atggataggg cctgtcaggc caagcttggg
     6961 gatcaattct ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     7021 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     7081 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     7141 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     7201 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctgggccta ctagatagcg
     7261 gaccccttac cgaaacatcg ccgcattctg cagaggagtc gaacttagtg tatgtaaact
     7321 tctgacccac tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa
     7381 ttgttggaaa aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac
     7441 tattgtttgt taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac
     7501 ttaagtgtat gtaaacttcc gacttcaact gtatagggat cc
//
