LOCUS       Exported                7145 bp DNA     circular SYN 02-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGAPDH-EGFP-NEO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2024 from SnapGene 5.3.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7145
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..744
                     /label=GAPDH promoter
     primer_bind     703..729
                     /label=seq
     CDS             761..781
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             782..1807
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             1814..2530
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     rep_origin      2733..3188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3346..3643
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3513..3648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    3647..3829
                     /label=TBP promoter
     CDS             3911..4930
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     polyA_signal    4994..5042
                     /note="synthetic polyadenylation signal"
     promoter        5348..5452
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5453..6313
                     /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      6484..7072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atatgcaacc ctgtgacgag agacagggag tgctgttaga gaagagcgca cattcagcgc
       61 tcaagtggtc ttgcgtttgg tgtcagcacg aggtgcgcca gacccaggtc gcacgcaggc
      121 ccctcagcgc ggtcgccgtg tgttggtatt gtgtgcctga gtgatgcgga atgcgttctg
      181 cttgtaggcg tacgagaccg acgccaggcg gcctgtcgtg ctgggctcgt ccgcggcagg
      241 ctctgtaggg ctggcgcgga gtgggcgatc gtcctcccgc cgcgaggaaa cagcaacgga
      301 caggcttcat tgccacctca tcgccgcttt cgcgctcgag gccagggcgt ggcgagcagg
      361 taagaatctg agaagttgaa ttgtgaatta acacaaagaa gacgaacgaa cccgccgtgg
      421 tatggaagaa ggccacctca accaatctca agatcacctc ctaatcgtcc aacccggcct
      481 acaatcgcgc acatcagcac tcgatctcgc acttcggtcc gtttggccct cgagccaaag
      541 ccaagcagtt ttccaggcac cgttgaagtg cgaatagatg aaatttccgg tctaccccaa
      601 tgacatggcg ttttgtcaca acacatggcg ctgtgctggt ggtctcggtg gggcctattt
      661 atttggagtt gtttgcttgc tctctttcat ctttgtcttc ctacacagca gcaaacacca
      721 ctctctcact aagaagatct tccatatgac tagtatggcc ccaaagaaga agcggaaggt
      781 ctccaattta ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga
      841 tgaggttcgc aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac
      901 ctggaaaatg cttctgtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg
      961 gaaatggttt cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg
     1021 cggtctggca gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg
     1081 gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg
     1141 aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga
     1201 tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct
     1261 ggcatttctg gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag
     1321 ggttaaagat atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa
     1381 aacgctggtt agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt
     1441 cgagcgatgg atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg
     1501 ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct
     1561 ggaagggatt tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca
     1621 gagatacctg gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg
     1681 cgctggagtt tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt
     1741 catgaactat atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga
     1801 tggcgatact agtatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1861 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1921 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1981 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2041 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2101 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2161 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2221 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2281 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2341 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2401 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2461 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2521 gctgtacaag tctagataag tcgacccaac aacaacaatt gcattcattt tatgtttcag
     2581 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     2641 atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2701 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     2761 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2821 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2881 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2941 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3001 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3061 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3121 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3181 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3241 cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta
     3301 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3421 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3481 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3541 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3601 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggatca aacgacgcct
     3661 tgcaacaagc tgagaaaaaa ccaggatcgg caaggaagga ttttcaacgg aaacgttggc
     3721 atcaccgggt ataaaaaggg gccaattttt ttgttgattt gttgcgcgaa ttcttgcttt
     3781 cggcatcgaa ttcaagggag aaggagtcga ttcacacata caacaagatc ctaggctttt
     3841 gcaaaaagct tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgga
     3901 tagatccgga aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     3961 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     4021 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     4081 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     4141 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     4201 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     4261 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     4321 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     4381 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     4441 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     4501 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4561 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4621 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4681 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4741 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     4801 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     4861 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     4921 aaaggaatag gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4981 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     5041 tgaatcgata gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc
     5101 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5161 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5221 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5281 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5341 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5401 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5461 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5521 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5581 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5641 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5701 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5761 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5821 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5881 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5941 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6001 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6061 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6121 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6181 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6241 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6301 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6361 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6421 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6481 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6541 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6601 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6721 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6781 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6841 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6901 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6961 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7021 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7081 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca
     7141 gatct
//
