LOCUS       62056_2465        6652 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 6652)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     protein_bind    complement(1434..1467)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1544..1577)
                     /label=loxP511
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             complement(1629..2330)
                     /codon_start=1
                     /label=dTomato
                     /note="dimeric variant of DsRed fluorescent protein (Shaner
                     et al., 2004)"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     protein_bind    2353..2386
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2779..3367
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3399..3875
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3915..4055
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4130..4585
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4867..4971
                     /label=AmpR promoter
     CDS             4972..5829
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6003..6591
                     /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 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagatcc ggaataactt
     1441 cgtatagcat acattatacg aagttatgca gaatggtagc tggattgtag tcgacgtgtg
     1501 ttttgagact ataaagctgc tattagcaat atgaaacctc ttaataactt cgtatagtat
     1561 aaattatacg aagttataag ccttgttaac gcgcggtgac cgctagcact agttactagg
     1621 atccgttact tgtacagctc gtccatgccg tacaggaaca ggtggtggcg gccctcggag
     1681 cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc cagcttggtg
     1741 tccacgtagt agtagccggg cagttgcacg ggcttcttgg ccatgtagat ggtcttgaac
     1801 tccaccaggt agtggccgcc gtccttcagc ttcagggcct ggtggatctc gcccttcagc
     1861 acgccgtcgc gggggtacag gcgctcggtg gaggcctccc agcccatggt cttcttctgc
     1921 attacggggc cgtcgggggg gaagttggtg ccgcgcatct tcaccttgta gatcagcgtg
     1981 ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac cgccgtcctc gaagttcatc
     2041 acgcgctccc acttgaagcc ctcggggaag gacagcttct tgtaatcggg gatgtcggcg
     2101 gggtgcttca cgtacgcctt ggagccgtac atgaactggg gggacaggat gtcccaggcg
     2161 aagggcaggg ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg
     2221 cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcatggagcc ctccatgcgc
     2281 accttgaagc gcatgaactc tttgatgacc tcctcgccct tgctcaccat ggtggcggtg
     2341 gcgaccggta gcataacttc gtataatgta tgctatacga agttatttgc cttaacccag
     2401 aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa tttatactat
     2461 acgaagttat aattacttta cagttagggt gagtttcctt ttgtgctgtt ttttaaaata
     2521 ataatttagt atttgtatct cttatagaaa tccaagccta tcatgtaaaa tgtagctagt
     2581 attaaaaaga acagattatc tgtcttttat cgcacattaa gcctctatag ttactaggaa
     2641 atattatatg caaattaacc ggggcagggg agtagccgag cttctcccac aagtctgtgc
     2701 gagggggccg gcgcgggcct agagatggcg gcgtcggatc tctagagcgg gcccgaattc
     2761 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2821 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2881 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2941 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3001 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3061 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3121 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3181 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     3241 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3301 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3361 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     3421 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     3481 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     3541 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     3601 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     3661 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     3721 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     3781 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     3841 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     3901 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3961 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4021 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     4081 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4141 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4201 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4261 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4321 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     4381 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4441 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     4501 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4561 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     4621 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4681 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4741 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     4801 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4921 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4981 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5041 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5101 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5161 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5221 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5281 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5341 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5401 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5461 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5521 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5581 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5641 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5701 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5761 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5821 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5881 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
