LOCUS       62056_8860        6505 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6505)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        430..1592
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1633..2349
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2374..2597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2624..3124
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3141..3941
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    complement(3951..4072)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(4201..4234)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4612..4716
                     /label=AmpR promoter
     CDS             4717..5574
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5748..6336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctagtcgagt aaagcgcaaa tcttttttaa ccctagaaag atagtctgcg taaaattgac
       61 gcatgcattc ttgaaatatc gctctctctt tctaaatagc gcgaatccgt cgctgtgcat
      121 ttaggacatc tcagtcgccg cttggagctc ccgtgaggcg tgtttgtcaa tgcggtaagt
      181 gtcactgatt ttgaactata acgaccgcgt gagtcaaaat gacgcatgat tatcttttac
      241 gtgactttta agatttaact catacgataa ttatattgtt atttcatgtt ctacttacgt
      301 gataacttat tatatatata ttttcttgtt atagatatcg tgactcatca cactggcggc
      361 cgcataactt cgtatagcat acattatacg aagttatcag ccatcgatgc cgatatccta
      421 gcttcgtgag gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      481 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
      541 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      601 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      661 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
      721 ccttgaatta cttccacctg gctccagtac gtgattcttg atcccgagct ggagccaggg
      781 gcgggccttg cgctttagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
      841 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      901 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      961 gtcttgtaaa tgcgggccag gatctgcaca ctggtatttc ggtttttggg cccgcggccg
     1021 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
     1081 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
     1141 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
     1201 gagcggaaag atggccgctt cccggccctg ctccaggggg ctcaaaatgg aggacgcggc
     1261 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
     1321 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
     1381 ggagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1441 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcgt
     1501 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1561 aaagtttttt tcttccattt caggtgtcgt gaacacgtgg tcgcggccat ctagagcaac
     1621 tagtgggcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1681 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1741 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1801 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1861 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1921 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1981 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2041 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2101 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2161 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2221 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2281 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2341 ctgtacaagt aaggatccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2401 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2461 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2521 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2581 atgcggtggg ctctatgaag cttaaatccc ggcgcgccct accgggtagg ggaggcgctt
     2641 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     2701 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     2761 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     2821 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     2881 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     2941 cagctttggc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     3001 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggaa gcccggcatt
     3061 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     3121 ttcgacctgc agccgctagc atgggatcgg ccattgaaca agatggattg cacgcaggtt
     3181 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3241 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3301 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3361 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3421 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3481 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3541 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3601 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3661 tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg
     3721 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3781 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3841 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3901 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaagatct taagatacat
     3961 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4021 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4081 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4141 gtaaaacctc tacaaatgtg gtatggctga ttatgatccg gctgcctcgc gccatatgtg
     4201 ataacttcgt atagcataca ttatacgaag ttatgaaaag ttttgttact ttatagaaga
     4261 aattttgagt ttttgttttt ttttaataaa taaataaaca taaataaatt gtttgttgaa
     4321 tttattatta gtatgtaagt gtaaatataa taaaacttaa tatctattca aattaataaa
     4381 taaacctcga tatacagacc gataaaacac atgcgtcaat tttacgcatg attatcttta
     4441 acgtacgtca caatatgatt atctttctag ggttaaataa tagtttctaa tttttttatt
     4501 attcagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4561 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4621 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4681 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4741 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4801 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4861 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4921 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     4981 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5041 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5101 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5161 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5221 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5281 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5341 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5401 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5461 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5521 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5581 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5641 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5701 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5761 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5821 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5881 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5941 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6001 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6061 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6121 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6181 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6241 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6301 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6361 cggttcctgg ccttttgctg gccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     6421 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     6481 gggaaggcca tccagcctcg cgtcg
//
