LOCUS       Exported                9583 bp ds-DNA     circular SYN 26-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9583)
  AUTHORS   molly
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 9, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9583
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     misc_feature    395..1626
                     /label=CAG promotor
     misc_feature    1744..5844
                     /label=IGF1R
     CDS             5863..6579
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_site      6672..6727
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(7086..7102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7110..7126
                     /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(7134..7164)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7179..7200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7259..7454
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7305..7440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7460..7594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(7833..8421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8592..9452)
                     /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(9453..9557)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cgggggcgag
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaagaatt cgctagcgcc
     1741 accatgaagt ctggctccgg aggagggtcc ccgacctcgc tgtgggggct cctgtttctc
     1801 tccgccgcgc tctcgctctg gccgacgagt ggagaaatct gcgggccagg catcgacatc
     1861 cgcaacgact atcagcagct gaagcgcctg gagaactgca cggtgatcga gggctacctc
     1921 cacatcctgc tcatctccaa ggccgaggac taccgcagct accgcttccc caagctcacg
     1981 gtcattaccg agtacttgct gctgttccga gtggctggcc tcgagagcct cggagacctc
     2041 ttccccaacc tcacggtcat ccgcggctgg aaactcttct acaactacgc cctggtcatc
     2101 ttcgagatga ccaatctcaa ggatattggg ctttacaacc tgaggaacat tactcggggg
     2161 gccatcagga ttgagaaaaa tgctgacctc tgttacctct ccactgtgga ctggtccctg
     2221 atcctggatg cggtgtccaa taactacatt gtggggaata agcccccaaa ggaatgtggg
     2281 gacctgtgtc cagggaccat ggaggagaag ccgatgtgtg agaagaccac catcaacaat
     2341 gagtacaact accgctgctg gaccacaaac cgctgccaga aaatgtgccc aagcacgtgt
     2401 gggaagcggg cgtgcaccga gaacaatgag tgctgccacc ccgagtgcct gggcagctgc
     2461 agcgcgcctg acaacgacac ggcctgtgta gcttgccgcc actactacta tgccggtgtc
     2521 tgtgtgcctg cctgcccgcc caacacctac aggtttgagg gctggcgctg tgtggaccgt
     2581 gacttctgcg ccaacatcct cagcgccgag agcagcgact ccgaggggtt tgtgatccac
     2641 gacggcgagt gcatgcagga gtgcccctcg ggcttcatcc gcaacggcag ccagagcatg
     2701 tactgcatcc cttgtgaagg tccttgcccg aaggtctgtg aggaagaaaa gaaaacaaag
     2761 accattgatt ctgttacttc tgctcagatg ctccaaggat gcaccatctt caagggcaat
     2821 ttgctcatta acatccgacg ggggaataac attgcttcag agctggagaa cttcatgggg
     2881 ctcatcgagg tggtgacggg ctacgtgaag atccgccatt ctcatgcctt ggtctccttg
     2941 tccttcctaa aaaaccttcg cctcatccta ggagaggagc agctagaagg gaattactcc
     3001 ttctacgtcc tcgacaacca gaacttgcag caactgtggg actgggacca ccgcaacctg
     3061 accatcaaag cagggaaaat gtactttgct ttcaatccca aattatgtgt ttccgaaatt
     3121 taccgcatgg aggaagtgac ggggactaaa gggcgccaaa gcaaagggga cataaacacc
     3181 aggaacaacg gggagagagc ctcctgtgaa agtgacgtcc tgcatttcac ctccaccacc
     3241 acgtcgaaga atcgcatcat cataacctgg caccggtacc ggccccctga ctacagggat
     3301 ctcatcagct tcaccgttta ctacaaggaa gcacccttta agaatgtcac agagtatgat
     3361 gggcaggatg cctgcggctc caacagctgg aacatggtgg acgtggacct cccgcccaac
     3421 aaggacgtgg agcccggcat cttactacat gggctgaagc cctggactca gtacgccgtt
     3481 tacgtcaagg ctgtgaccct caccatggtg gagaacgacc atatccgtgg ggccaagagt
     3541 gagatcttgt acattcgcac caatgcttca gttccttcca ttcccttgga cgttctttca
     3601 gcatcgaact cctcttctca gttaatcgtg aagtggaacc ctccctctct gcccaacggc
     3661 aacctgagtt actacattgt gcgctggcag cggcagcctc aggacggcta cctttaccgg
     3721 cacaattact gctccaaaga caaaatcccc atcaggaagt atgccgacgg caccatcgac
     3781 attgaggagg tcacagagaa ccccaagact gaggtgtgtg gtggggagaa agggccttgc
     3841 tgcgcctgcc ccaaaactga agccgagaag caggccgaga aggaggaggc tgaataccgc
     3901 aaagtctttg agaatttcct gcacaactcc atcttcgtgc ccagacctga aaggaagcgg
     3961 agagatgtca tgcaagtggc caacaccacc atgtccagcc gaagcaggaa caccacggcc
     4021 gcagacacct acaacatcac cgacccggaa gagctggaga cagagtaccc tttctttgag
     4081 agcagagtgg ataacaagga gagaactgtc atttctaacc ttcggccttt cacattgtac
     4141 cgcatcgata tccacagctg caaccacgag gctgagaagc tgggctgcag cgcctccaac
     4201 ttcgtctttg caaggactat gcccgcagaa ggagcagatg acattcctgg gccagtgacc
     4261 tgggagccaa ggcctgaaaa ctccatcttt ttaaagtggc cggaacctga gaatcccaat
     4321 ggattgattc taatgtatga aataaaatac ggatcacaag ttgaggatca gcgagaatgt
     4381 gtgtccagac aggaatacag gaagtatgga ggggccaagc taaaccggct aaacccgggg
     4441 aactacacag cccggattca ggccacatct ctctctggga atgggtcgtg gacagatcct
     4501 gtgttcttct atgtccaggc caaaacagga tatgaaaact tcatccatct gatcatcgct
     4561 ctgcccgtcg ctgtcctgtt gatcgtggga gggttggtga ttatgctgta cgtcttccat
     4621 agaaagagaa ataacagcag gctggggaat ggagtgctgt atgcctctgt gaacccggag
     4681 tacttcagcg ctgctgatgt gtacgttcct gatgagtggg aggtggctcg ggagaagatc
     4741 accatgagcc gggaacttgg gcaggggtcg tttgggatgg tctatgaagg agttgccaag
     4801 ggtgtggtga aagatgaacc tgaaaccaga gtggccatta aaacagtgaa cgaggccgca
     4861 agcatgcgtg agaggattga gtttctcaac gaagcttctg tgatgaagga gttcaattgt
     4921 caccatgtgg tgcgattgct gggtgtggtg tcccaaggcc agccaacact ggtcatcatg
     4981 gaactgatga cacggggcga tctcaaaagt tatctccggt ctctgaggcc agaaatggag
     5041 aataatccag tcctagcacc tccaagcctg agcaagatga ttcagatggc cggagagatt
     5101 gcagacggca tggcatacct caacgccaat aagttcgtcc acagagacct tgctgcccgg
     5161 aattgcatgg tagccgaaga tttcacagtc aaaatcggag attttggtat gacgcgagat
     5221 atctatgaga cagactatta ccggaaagga gggaaagggc tgctgcccgt gcgctggatg
     5281 tctcctgagt ccctcaagga tggagtcttc accacttact cggacgtctg gtccttcggg
     5341 gtcgtcctct gggagatcgc cacactggcc gagcagccct accagggctt gtccaacgag
     5401 caagtccttc gcttcgtcat ggagggcggc cttctggaca agccagacaa ctgtcctgac
     5461 atgctgtttg aactgatgcg catgtgctgg cagtataacc ccaagatgag gccttccttc
     5521 ctggagatca tcagcagcat caaagaggag atggagcctg gcttccggga ggtctccttc
     5581 tactacagcg aggagaacaa gctgcccgag ccggaggagc tggacctgga gccagagaac
     5641 atggagagcg tccccctgga cccctcggcc tcctcgtcct ccctgccact gcccgacaga
     5701 cactcaggac acaaggccga gaacggcccc ggccctgggg tgctggtcct ccgcgccagc
     5761 ttcgacgaga gacagcctta cgcccacatg aacgggggcc gcaagaacga gcgggccttg
     5821 ccgctgcccc agtcttcgac ctgcggcgga agcggcggat ccgtgagcaa gggcgaggag
     5881 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     5941 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     6001 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     6061 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     6121 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     6181 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     6241 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     6301 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     6361 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     6421 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     6481 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     6541 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagagtc
     6601 gacggatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     6661 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     6721 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     6781 ggtttagagt ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa
     6841 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     6901 tgacttgagg ttagattttt ttttatattt tgtttgtgtt atttttttct ttaacatccc
     6961 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     7021 tcatagctgt ccctcttctc ttatgaagat ccctcgacct gcagcccaag cttggcgtaa
     7081 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7141 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7201 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     7261 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     7321 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7381 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7441 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     7501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7561 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     7621 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     8041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     8281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8461 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8521 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8581 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8641 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8701 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8761 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8821 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8881 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8941 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9001 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9061 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9121 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9181 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9241 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9301 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9361 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9421 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9481 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9541 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
