LOCUS       Exported                7326 bp DNA     circular SYN 01-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7326)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7326
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=ITR???
     misc_feature    183..608
                     /label=hsyn
     misc_feature    609..620
                     /label=kozak
     protein_bind    complement(627..660)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(711..744)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     CDS             complement(751..2325)
                     /codon_start=1
                     /label=N2cG
                     /translation="MVPQVLLFVLLLGFSLCFGKFPIYTIPDELGPWSPIDIHHLSCPN
                     NLVVEDEGCTNLSEFSYMELKVGYISAIKVNGFTCTGVVTEAETYTNFVGYVTTTFKRK
                     HFRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVRTTKESLIIISPSVTDLDP
                     YDKSLHSRVFPGGKCSGITVSSTYCSTNHDYTIWMPENPRPRTPCDIFTNSRGKRASNG
                     NKTCGFVDERGLYKSLKGACRLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHD
                     FRSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKT
                     LMEADAHYKSVRTWNEIIPSKGCLKVGGRCHPHVNGVFFNGIILGPDDHVLIPEMQSSL
                     LQQHMELLESSVIPLMHPLADPSTVFKEGDEAEDFVEVHLPDVYKQISGVDLGLPNWGK
                     YVLMTAGAMIGLVLIFSLMTWCRRANRPESKQRSFGGTGGNVSVTSQSGKVIPSWESYK
                     SGGEIRL"
     CDS             complement(2326..2382)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2392..2398)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="YK"
     CDS             complement(2399..3108)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDE"
     CDS             complement(3109..3165)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(3174..3643)
                     /codon_start=2
                     /label=TVA
                     /translation="ARLLPALLLLLLPGNVTGNGSGNGSLSRCPPGQFRCSEPPGAHGE
                     CYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPAPGRALPARNHGRMWM
                     LITAVLLCCLVAVGGIAAWGKSKAKSRSDIFSLESASKELLVPDKSQADLFS"
     primer_bind     3316..3339
                     /label=N2c_Fcorrect
     misc_feature    3644..3647
                     /label=?
     protein_bind    3648..3681
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    3732..3765
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    3767..4355
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3834..3854)
                     /label=Rcommon
     CDS             complement(4238..4249)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4363..4587
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4589..4729
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4804..5259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5541..5645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5646..6506
                     /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      6677..7265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcagc cgccaccatg gttaatataa cttcgtataa tgtatgctat acgaagttat
      661 gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta ataacttcgt
      721 ataaagtatc ctatacgaag ttatggtacc ttacagtctg atctcacctc cactcttata
      781 tgattcccat gaaggtatga cttttccgct ttgggaagtg actgacacat tcccccctgt
      841 ccctccaaaa ctgcgttgtt tcgattctgg tcgattggct cttctgcacc atgtcattag
      901 ggaaaatatc aacaccaggc caatcatggc ccctgcagtc atcaatacat actttcccca
      961 gttcgggaga cccaggtcaa cccctgagat ctgtttgtac acatcgggga ggtgaacttc
     1021 aacaaaatcc tcagcctcat caccttcttt gaaaactgta gaagggtctg ccagggggtg
     1081 catcaggggg ataactgaag attccaacaa ctccatatgt tgctggagga gggatgattg
     1141 catctctggg attaggacat ggtcgtcagg ccctaatatt ataccattga aaaacacccc
     1201 gttcacatga ggatggcacc ttcctccaac tttcaaacac ccttttgagg ggatgatctc
     1261 attccaggtc cggactgact tgtagtgagc atcagcctcc atcaaggttt tgttgaatat
     1321 ggtatatgct tttccaaacc ctgggacaag ttttctcagg tgactgagac gtctgaaact
     1381 tactgacttg gtggtcatga tggactctaa tgcatccaga cattcctctc ttttcttgac
     1441 taactcctcc acaacgagat gctcaatctc gtctgagcga aagtcgtgca aattcaccaa
     1501 ctgatctgga gggcaccatt tggtctcatc tgatgtttgc atcgcgaccc atgttccatc
     1561 cataagtcta agtccaagaa ctccacataa cttgagcctg catgctcctt ttagagactt
     1621 atacaggcct ctttcatcca caaagccgca agtcttgttc ccgttggatg ctctcttccc
     1681 tctgctattg gtaaaaatgt cacaaggtgt ccttggtctc ggattctcgg gcatccaaat
     1741 ggtgtaatca tggttagttg agcagtaggt agaggacacc gttattcctg agcactttcc
     1801 gccagggaag acccttgagt gaagggattt gtcatatggg tccaaatctg tcacacttgg
     1861 ggatatgata atgagggact ctttggtggt tcttacagtt cgaagccagt ggtagtcggg
     1921 gtatggattg tgtagggact cttcatatct ggggtcaccg gccatcttcc agttatacgc
     1981 ggctctacat gcgtctgggg tggggcggaa atgctttctc ttgaatgtgg ttgtgacata
     2041 accaacaaag ttggtgtagg tctctgcctc tgtcacaaca cctgtgcaag tgaacccgtt
     2101 cactttgatg gctgagatgt atcccacttt gagttccatg taggagaact cggacaggtt
     2161 ggtacatcct tcatcctcca caaccaggtt atttggacag ctgagatggt gtatgtcaat
     2221 agggctccag ggaccaagtt cgtctggtat cgtgtaaatg gggaacttcc cgaaacacaa
     2281 cgaaaaaccc agaaggagta caaacaaaag aacctgagga accataggtc cagggttctc
     2341 ctccacgtct ccagcctgct tcagcaggct gaagttagta gctccgcttc ccttgtacag
     2401 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     2461 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
     2521 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     2581 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     2641 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     2701 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     2761 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     2821 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     2881 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     2941 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     3001 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     3061 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg gtccagggtt
     3121 ctcctccacg tctccagcct gcttcagcag gctgaagtta gtagctccgg atccggagaa
     3181 caagtctgcc tggctcttgt caggcaccag cagctccttg gatgcgcttt caagactgaa
     3241 gatgtcagac ctgctttttg ctttggactt cccccatgca gcgataccac ccacagctac
     3301 caggcagcac aggagcactg cagtgatcag catccacatg cggccgtgat tcctggctgg
     3361 cagagcacgt ccaggagcag gggcagtggg agcctctgtg ccgttgtccg tgggcaccgc
     3421 ggggatcgcg ctggtcccgc agccccactc gtcccgcccg tcgtcgcagt cggggtgtcc
     3481 gtcgcacagc cagtcctgcg ggtaacactc cccgtgggca ccgggcggct ccgagcagcg
     3541 gaactgaccg ggggggcaac gggacaaaga accgttaccg gacccgttac cggtcacgtt
     3601 accgggcagc agcagcagca gcagcgcggg cagcagccgc gcacctaata acttcgtata
     3661 gcatacatta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     3721 tggtgcaaag aataacttcg tataggatac tttatacgaa gttatgaatc aacctctgga
     3781 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3841 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3901 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3961 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4021 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4081 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4141 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4201 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4261 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4321 gacgagtcgg atctcccttt gggccgcctc cccgcctgga tcctgtgcct tctagttgcc
     4381 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4441 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4501 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4561 atgctgggga tgcggtgggc tctatggcag gaacccctag tgatggagtt ggccactccc
     4621 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4681 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4741 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4801 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4981 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     5161 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5221 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5281 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5341 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5401 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5461 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5521 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5581 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5641 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5701 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5761 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5821 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5881 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5941 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6001 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6061 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6121 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6181 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6241 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6301 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6361 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6421 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6481 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6541 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6601 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6661 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6721 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6781 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6841 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6901 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6961 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7021 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7081 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7141 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7201 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7261 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7321 acatgt
//
